pinpoint colorimetric developer

ErbB2driven breast cancer cell invasion depends on a complex signaling network activating myeloid zinc finger1dependent cathepsin B expression. 245 0 R null null null 246 0 R null null null 247 0 R null Screenshot: livebar opened / hidden in a mobile (vertical) screen, Screenshot: livebar opened on a wide screen. Briefly, 200ng of qualified RNA was added to the assay wells provided by the kit. /Keywords () Previous studies have demonstrated that ARL5B overexpression facilitated lysosome motility, resulting in lysosome dispersion and accumulation at the cell periphery [38, 39]. 145 0 R 146 0 R null null 236 0 R null null] /C2_0 45 0 R The closure of the gap was imaged at designated time intervals under 100 field, and the wound closure speed was evaluated using the ImageJ software. It is a customizable product that combines content and services in a geographical setting. Transfection was conducted using Lipofectamine 3000 (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA) according to the manufacturer's instructions. Aerosol)WDP-217 is an aerosol water leak detector/color change developer, which when sprayed on the points of suspected leaks (joints, welds, W-15A-M Water Leak Detection Monitors (4" x 4.25", pkg of 12)W-15A-M PinPoint Colormetric Paper for Water Leak Detection (4" x 4.25" adhesive back, pkg of 12 sheets to a carton). THW and PS designed and funded the study. NOTE: THIS PRODUCT CAN ONLY BE SHIPPED FEDEX GROUND SERVICES. >> A scatter plot of the relative correlation between FTO and ARL5B expression was performed using the R software. 250 0 R null null null 251 0 R null null null 252 0 R null 102 0 R 103 0 R 104 0 R 105 0 R 106 0 R 107 0 R 108 0 R 109 0 R 110 0 R 111 0 R /Parent 4 0 R /StructParents 1 Written informed consent was obtained from all patients. Either your web browser doesn't support Javascript or it is currently turned off. /Marked true In summary, these data indicate that FTO could accelerate the migration and invasion of HER2positive breast cancer cells via the FTO/miR181b3p/ARL5B signaling pathway (Figure10). [13] showed that FTO reduced the transcription of ankyrin repeat and SOCS box containing 2 (ASB2) and retinoic acid receptor alpha (RARA) by serving as an m6A demethylase, promoting proliferation and inhibiting apoptosis in acute myeloid leukemia. E Wound healing assay of SKBR3 and MDAMB453 cells pretreated with gradient concentrations of MA2 for 48 h. DMSO was used as solvent control. H Transwell invasion assay of SKBR3 and MDAMB453 cells transfected with FTO plasmid. Su etal. 4 Department of Basic Medicine, Zhongshan School of Medicine, Sun Yatsen University, Guangzhou Guangdong, 510080 P. R. China. Significant differences were calculated by twotailed Student's ttest or oneway analysis of variance (oneway ANOVA). My family immigrated to the USA in the late 60s. SHZ and KWZ provided supervision and suggestions for the implementation of the study. Due to the m6A demethylase function of FTO, we found that m6A was irregularly downregulated in HER2positive breast cancer tissues (Figure1C). A livemap contains several high-level, functional components that interact with each other. Further studies are urged to fully unveil the comprehensive role of FTO in breast cancer. Plasmid clones expressing FTO (oeFTO), ARL5B (oeARL5B), or empty vector (oeVec) were purchased from Sino Biological Inc. (Beijing, China). >> 130 0 R 131 0 R 132 0 R 133 0 R 134 0 R] By wound healing assay in SKBR3 cells and Transwell migration assay in MDAMB453 cells, cell migration was decreased upon FTO knockdown but accelerated upon FTO overexpression (Figure2C2D). Abbreviations: miRNC, negative control of miR181b3p mimics; miRNC2, negative control of miR181b3p inhibitors; ARL5B, ADP ribosylation factor like GTPase 5B; RAP2C, Rasrelated protein Rap2c; CLTA, clathrin light chain A; PCMTD1, proteinLisoaspartate (Daspartate) Omethyltransferase domain containing 1; RIMKLB, ribosomal modification protein rimK like family member B; SOWAHB, sosondowah ankyrin repeat domain family member B; other abbreviations as in Figure2. /Im2 40 0 R Ltd. is ISO 9001 Certified Company.Gas Leak Detection Sprays is an aerosol product that reveals the tiniest gas leaks in all sealed systems. 3 0 obj The preview is a media on the background (image thumbnail or icon) and a customizable subset of the pinpoints content in the livebar. Welcome to Iggy Garcia, The Naked Shaman Podcast, where amazing things happen. The livebar is highly customizable both in terms of User Interface (layout) and User Experience (behaviour). Screenshot - a livemap with third-party tiles in the background. stream /Type /Page This observation implied the intricate involvement of FTO in different breast cancer molecular subtypes, which is worthy of further exploration. The fully fledged editing tool allows you to define the parameters of your background style. Moreover, FTO promoted cell invasion and migration in vitro. Correlation analysis revealed that the expression levels of FTO and ARL5B were positively correlated (Figure7D), and the dual luciferase reporter assay confirmed that miR181b3p bound directly to the ARL5B 3UTR (Figure7E7F). Statistical analyses were performed using IBM SPSS Statistics version 22.0 (SPSS Inc., Chicago, IL, USA) and GraphPad Prism 7.0 (GraphPad Software Inc., La Jolla, CA, USA). A 10 L sterile pipette tip was used to scratch the cell layer and form a wound. Gas Leak Detection Sprays is an aerosol product that reveals the tiniest gas leaks in all sealed systems. 79 0 R null null 160 0 R null null null 161 0 R null null] /Contents [53 0 R 54 0 R 55 0 R] Received 2020 Jan 4; Revised 2020 Apr 22; Accepted 2020 Jun 27. /Manager (Publications) Correspondingly, FTO inhibition by siRNA or MA2 was found to impair cell invasion (Figure2F and2G), while the results were reversed when FTO was overexpressed (Figure2H). /TT3 34 0 R Conference Proceedings /MarkInfo << /S /Sect RNA sequencing (RNAseq) was conducted to decipher the downstream targets of FTO. This preview can be hidden depending on the amount of pinpoints in the visible (active) area of the map background. 208 0 R 209 0 R 210 0 R 211 0 R 212 0 R 213 0 R 214 0 R 215 0 R 216 0 R 217 0 R Our leak detection spray does not form harmful compounds with the following gases: For all compressed air, natural gas or liquid gas piping jobs, Detecting leakages on fittings, threaded joints, and connections, Detection of leaks in high-pressure pipes, valves, and compressed air cylinders. /P 5 0 R /MediaBox [0 0 595.32 842.04] In the present study, we discovered that FTO was highly expressed in HER2positive breast cancer cells and tissues. 1 Place Francis Ponge All these functions are offered in a feature rich and WYSIWYG interface, allowing a powerful and seamless way to create rich local information experiences tailored to your audiences needs. Abbreviations: ARL5B, ADP ribosylation factor like GTPase 5B; siARL5B, ARL5B siRNA; oeARL5B, ARL5B clone plasmid; other abbreviations as in Figure2, ARL5B plays an oncogenic role in HER2positive breast cancer cells. Pinpoints are geolocalized pieces of content present in the livemap. endobj Overexpressing ARL5B after MA2 treatment also rescued the inhibitory effect of MA2 (Figure8E and8F). Pan H, Gray R, Braybrooke J, Davies C, Taylor C, McGale P, etal. 2 /ParentTreeNextKey 7 /Rotate 0 Lysosomal trafficking, antigen presentation, and microbial killing are controlled by the Arflike GTPase Arl8b, Arl8 and SKIP act together to link lysosomes to kinesin1, The Arffamily protein, Arl8b, is involved in the spatial distribution of lysosomes, The small G protein Arl8 contributes to lysosomal function and longrange axonal transport in Drosophila. Zhou K, Sun P, Zhang Y, You X, Li P, Wang T. Estrogen stimulated migration and invasion of estrogen receptornegative breast cancer cells involves an ezrindependent crosstalk between G proteincoupled receptor 30 and estrogen receptor beta signaling, Overexpression of the fat mass and obesity associated gene (FTO) in breast cancer and its clinical implications. Furthermore, m6A quantification assay showed that inhibition of FTO increased m6A levels, while FTO overexpression decreased m6A abundance (Figure2I). /CropBox [0 0 595.32 842.04] /DropCap /Figure The authenticity of all cell lines was confirmed by short tandem repeat (STR) DNA profiling analysis. /F6 36 0 R /Parent 4 0 R Sold under the brand name of Spray Fill for more than 30 years. /TT3 35 0 R /Parent 4 0 R D:20100920202336 In the present study, miR181b3p inhibited the migration and invasion of HER2positive breast cancer cells by targeting ARL5B. Protein concentration was determined with the BCA protein assay kit (Thermo Fisher Scientific, Inc.). << All miR1813p mimics, inhibitors, and corresponding negative controls (miRNC, miRNC2) used for transfection were purchased from GenePharma Co., Ltd. (Shanghai, China). Steffan JJ, Williams BC, Welbourne T, Cardelli JA. << /TT1 33 0 R Li etal. The search engine is present natively on the livemap and provides the viewer with an additional ability to navigate the livemap and explore its content (pinpoints, events, lists). Knockdown and overexpression of FTO were carried out in SKBR3 and MDAMB453 cells, and corresponding changes in the expression of FTO were confirmed in both cell lines (Figure2A2B). Experiments were repeated independently at least three times. 1 Department of Physiology, Zhongshan School of Medicine, Sun Yatsen University, Guangzhou Guangdong, 510080 P. R. China. The optimal cutoff value of FTO (8.0) generated from the receiver operating characteristic (ROC) curve divided FTO expression into high or low. HGFinduced invasion by prostate tumor cells requires anterograde lysosome trafficking and activity of Na+H+ exchangers. To learn more about pinpoints, refer to entity description here. Dominissini D, MoshitchMoshkovitz S, Schwartz S, SalmonDivon M, Ungar L, Osenberg S, etal. /Font << Su R, Dong L, Li C, Nachtergaele S, Wunderlich M, Qing Y, etal. BC qRTPCR analysis of the expression of the six candidate molecules in SKBR3 and MDAMB453 cells with the transfection of miR181b3p mimics. The description and the details of the events are visible when the pinpoint is opened. /Kids [7 0 R 8 0 R 9 0 R 10 0 R 11 0 R 12 0 R 13 0 R] Rafn B, Nielsen CF, Andersen SH, Szyniarowski P, CorcelleTermeau E, Valo E, etal. << 14 0 obj /StructParents 3 C Relative m6A content of total RNA in paired HER2positive breast cancer tissues and adjacent nontumor tissues (n=12). 168 0 R 169 0 R 170 0 R 171 0 R 172 0 R 173 0 R 174 0 R 175 0 R 176 0 R 177 0 R 149 0 R 150 0 R 151 0 R 152 0 R null null 244 0 R null null null /Count 7 The contribution of FTO to the risk of various cancers shown in epidemiological studies [12], along with its aberrant expression in breast cancer, indicates the possible involvement of FTO in mediating the pathological development of breast cancer. Wound healing assay and Transwell assay were performed to determine the migration and invasion abilities of SKBR3 and MDA-MB453 cells with either knockdown or overexpression of FTO. We selected miR181b3p, the miRNA with the greatest change, to test the relationship between FTO and miRNAs in HER2positive breast cancer. Data are presented as the mean standard deviation. These results may imply the direct interaction between FTO and miR181b3p. /C2_0 31 0 R RNA sequencing (RNA-seq) was conducted to decipher the downstream targets of FTO. /ColorSpace << Tinghuai Wang, Email: nc.ude.usys.liam@htgnaw. 142 0 R 143 0 R 144 0 R 145 0 R 146 0 R 147 0 R 148 0 R 149 0 R 150 0 R 151 0 R Contact us. [23] searching for the possible consensus sequence motifs for m6A modification in m6A immunoprecipitated miRNAs, we succeeded in mapping the miR181b3p sequence to all the most discriminating motifs, and the mapped sequence was enriched in the center section. /TT0 32 0 R Abbreviations: FTO, fat mass and obesityassociated; HER2, human epidermal growth factor receptor 2; m6A, N6methyladenosine; HE, hematoxylin and eosin; IHC, immunohistochemistry; DFS, diseasefree survival; OS, overall survival; RFS, recurrencefree survival. /CS0 [/ICCBased 30 0 R] Wemap is a proud member of the OpenStreetMap community and our mapping data is synchronized with the OpenStreetMap dataset: this enables you to create your own style by choosing among numerous features and options including, among others: You have more specific needs? For the Transwell invasion assay, briefly, 5 1041 105 cells in 200 L serumfree DMEM were reseeded into the top of the insert of a Boyden chamber (Corning Inc., Corning, NY, USA) with 300g/mL Matrigel (BD Bioscience, San Jose, CA, USA), while 600 L medium with 10% FBS was loaded into the well below. Mechanistically, in SKBR3 and MDAMB453 cells, FTO accelerated cell migration and invasion by targeting ARL5B via miR181b3p downregulation. Schiefermeier N, Scheffler JM, de Araujo ME, Stasyk T, Yordanov T, Ebner HL, etal. >> /Font << The illumination of its regulatory mechanism will be helpful to clarify the clinical significance of miR181b3p in different circumstances [34]. background selection, within a catalogue of styles that are tailored for print (colorimetric adjustments, reduction of the geographic feature set to enhance readability on a printed material, etc. Abbreviations are the same as in Figure2, miR181b3p inhibits the protumor effects of FTO in HER2positive breast cancer cells. To explore potential targets of FTO, we performed RNAseq on FTOknockdown SKBR3 cells. Overall, ARL5B was identified as the downstream target of interest. Moreover, FTO promoted cell invasion and migration in vitro. /TT2 33 0 R Within Wemap Pro the Wemap Platform can be explored through a search engine that can be used to inspire and re-use content (pinpoints, lists, livemaps). /C2_0 45 0 R /Chart /Figure The binding site of miR181b3p to the ARL5B 3untranslated region (3UTR) was predicted using the miRNA databases mentioned above. Chua BA, Van Der Werf I, Jamieson C, Signer RAJ. /Type /Page Further studies are needed to fully unfold the clinical value of FTO, miR181b3p, and ARL5B in breast cancer. The background can also include some additional interactive layers (polygons, etc.). MicroRNAs (miRNAs) are a family of short noncoding singlestranded RNAs containing 1922 nucleotides that promote the degradation or translation inhibition of mRNA by targeting specific mRNAs and forming RNAinduced silencing complexes [19]. /Font << Gas Leak Detection Sprays can be used all types of plant, irrespective of the type of gas used.Several features are: We import Gas Leak Detection Sprays from FAREN Chemical Industries, Italy. /Rotate 0 The shaded area denoted the complementary bases and corresponding mutated nucleobases. All components are highly customizable, both in terms of User Interface and of feature-set. In addition, we observed inhibition of cell migration with MA2 treatment, and the inhibition effect was maximal at 80mol/L (Figure2E). Huang Y, Yan J, Li Q, Li J, Gong S, Zhou H, etal. 450) was measured using a microplate reader (SpectraMax M5 Molecular Devices, LLC, Sunnyvale, CA, USA) to calculate the m6A levels. /Parent 4 0 R Easy to clean off. D Representative IHC images of HER2 and FTO expression in tumor and nontumor tissues. /Author (European Space Agency) The manual / bulk creation of pinpoints from Wemap Pro offers a wide range of options to create rich content to be consumed from livemaps: media-rich pinpoints, events, html formatted descriptions, tags, synchronisation with social media accounts, and much more. or via an external application (app) that is synchronized with the Wemap platform. Therefore, 80mol/L MA2 was used for subsequent studies. Membranes were incubated with primary antibodies at 4C overnight and then secondary antibodies at room temperature for 2 h. After washing, signals were detected using a ChemiDoc imaging system (BioRad Laboratories, Inc.). Afterwards, the Venn diagram online tool (http://bioinformatics.psb.ugent.be/webtools/Venn/) was utilized to narrow down the scope into six target molecules. /StructParents 4 /Length 10 Ragvin A, Moro E, Fredman D, Navratilova P, Drivenes, Engstrm PG, etal. Lambert AW, Pattabiraman DR, Weinberg RA. For more details see livemap options and styling module. /Type /Page Colormetric detectors are rapid leak ABCD-311-18 Acid Base PiNPOiNT Colormetric Developer (12oz Aerosol [12 cans]) << Mechanistically, RNA-seq and further confirmation studies suggested that FTO up-regulated ARL5B by inhibiting miR-181b-3p. /Footnote /Note RNAseq analysis revealed that 19 miRNAs were significantly downregulated, while 10 miRNAs were significantly upregulated (Figure3A) the top six upregulated or downregulated miRNAs are listed in Figure3B. endstream >> >> On Wemap Pro a user will access its profile page to define or edit its personal information but also tailor her access rights, depending on the chosen Wemap subscription. Pinpoint utilizes only the most advanced electronic leak detection devices, including infrared thermal imaging, combined with proprietary techniques to pinpoint the exact location of your leak. In the present study, we confirmed that FTO could reduce the expression of miR181b3p, which inhibited the protumor effects of FTO. The results showed that FTO reinforced ARL5B expression, while miR181b3p inhibited ARL5B expression (Figure7A7C). /CS0 [/ICCBased 30 0 R] For more information, please visit: /TT2 52 0 R DE qRTPCR analysis of the expression of the six candidate molecules in SKBR3 and MDAMB453 cells with the transfection of miR181b3p inhibitors. /PageLayout /SinglePage Screenshot - Wemap Pro - explore the platform. Ltd. is ISO 9001 Certified Company. /ColorSpace << P value<0.05 was considered statistically significant. After 48 h of cotransfection, luciferase reporter assays were performed using the Dual Luciferase Reporter Assay System (Promega Corporation, Madison, WI, USA). As previously described [26], cultured cells were seeded into 24well plates at a density of 2 1054 105 cells per well. Cancer cells subcultured for less than 56 times were used in all experiments. From the water meter to anywhere in your home or building, including outside rain or moisture infiltration. Berulava T, Rahmann S, Rademacher K, KleinHitpass L, Horsthemke B. Ye S, Xu Y, Li J, Zheng S, Sun P, Wang T. Prognostic role of GPER/Ezrin in triple negative breast cancer is associated with menopausal status. stream uuid:e0c7ab79-8469-4935-9ee1-ca369aba8c50 We further verified that ARL5B also displayed carcinogenic activity in breast cancer cells.

Conclusion

Our work demonstrated the carcinogenic activity of FTO in promoting the invasion and migration of breast cancer cells via the FTO/miR-181b-3p/ARL5B signaling pathway. >L]"ZIa+~lEc' ${'8PBLO{. [44] detected that decreased lysosomal outward trafficking resulted in inhibited cancer cell invasion. /TT2 34 0 R AB The transfection efficiency of FTO siRNA or overexpression plasmid in SKBR3 and MDAMB453 cells confirmed by qRTPCR (A) and Western blotting (B). endobj stream << In the current study, we provide clinical and in vitro evidence that FTO, which is highly expressed in HER2positive breast cancer, could promote cell migration and invasion through the FTO/miR181b3p/ARL5B signaling pathway. /Type /Page /F8 38 0 R 1 [null null 162 0 R null null null 163 0 R null null null qRTPCR analysis confirmed that FTO inhibition using siRNA or MA2 resulted in significantly increased levels of miR181b3p, while FTO overexpression reduced miR181b3p expression (Figure4). The RTCGA. N6methyladenosine (m6A) modification, with methylated adenosine at the N6 position, is the most abundant internal modification of eukaryotic mRNA and noncoding RNA [7]. /StructParents 2 152 0 R 153 0 R 154 0 R 155 0 R] >> Moreover, based on the study by Berulava etal. >> /C2_0 31 0 R However, cell proliferation was not distinctly changed after FTO alteration (Supplementary Figure S1B). Screenshot - Airport livemap with multilevel management. endobj /TT1 52 0 R ADP-319-05 GAL PINPOINT COLORMETRIC- AMMONIA. Garg S, Sharma M, Ung C, Tuli A, Barral DC, Hava DL, etal. /Resources << /Type /Pages /Font << /TT0 32 0 R endobj /Rotate 0 /TextBox /Art 6 [null null 237 0 R null null null 238 0 R null null null /Outline /Span >> If youre curious about my background and how I came to do what I do, you can visit my about page. qRT-PCR, luciferase reporter assay, and Western blotting were employed to confirm the existence of the FTO/miR-181b-3p/ARL5B axis. /MediaBox [0 0 595.32 842.04] /Rotate 0 qRTPCR was performed using the SYBRGreen Master mix (Takara) in a CFX96 touch system (BioRad Laboratories, Inc., Hercules, CA, USA). Methyltransferase like 14 (METTL14) facilitates DiGeorge syndrome critical region 8 (DGCR8) recognition by adding m6A modification to primiR126, subsequently leading to the improvement of hepatic carcinoma [21]. Screenshot - Wemap Pro - livemaps management, Screenshot - Wemap Pro - livemap details management. The miR181 family has been clarified to be critical for cell cycle progression and differentiation [31]. All reads were mapped to human genome 19 using BWA tools and annotated by miRBase version 21 (https://www.mirbase.org) using miRDeep2 (https://www.mdc-berlin.de/8551903/en/). [27] discovered that FTO expression was significantly associated with clinicopathological characteristics of HER2positive breast cancer patients, which aligns with our current results. * P<0.05, ** P<0.01, *** P<0.001. /Endnote /Note Topology of the human and mouse m6A RNA methylomes revealed by m6Aseq. The followup deadline was September 20, 2019. If user has the access rights, the print module is available directly from Wemap Pro. 27 0 R 28 0 R 29 0 R] A Relative expression of FTO protein in breast cancer cell lines compared with the immortalized mammary epithelial cell line MCF10A. >> /C2_0 31 0 R The content on the livebar is always synchronized with the content displayed above the background to provide a consistent browsing experience. Screenshot: Livemap - web - mobile screen, Screenshot: livemap - web - desktop view - simplified chinese, Screenshot: livemap - mobile view - with itinerary, Screenshot: livemap - public screen (muse du Louvre). /Font << Experiments were terminated after scratch for 24 h. GH Transwell invasion assay in ARL5Bsilenced or overexpressing SKBR3 and MDAMB453 cells. endobj 16 0 obj B The top six upregulated or downregulated miRNAs. * P<0.05, ** P<0.01, *** P<0.001. In our present study, we failed to pinpoint the exact m6A modification site in miR181b3p by MeRIPseq. A methylated RNA immunoprecipitation sequencing (MeRIPseq) study showed that the steadystate levels of several miRNAs were affected by the knockdown of FTO and that a significant fraction of miRNAs contained m6A sites [23]. 30, Ground Floor, Ashoka Shopping Center LT Marg Mumbai - 400001, Maharashtra, India, No. The guided outward movement of lysosomes is considered to be a critical determinant of lysosomal function [42, 43]. The background component consists of a map made of tiles (vector or raster), hosted on Wemap servers and customizable (language, graphic styling, indoor mapping, etc.). 188 0 R 189 0 R 190 0 R 191 0 R 192 0 R 193 0 R 194 0 R 195 0 R 196 0 R 197 0 R 2 0 obj >> ARL5B is the target of miR181b3p and positively regulated by FTO. Experiments were terminated after seeding for 24 h. I Relative m6A content of total RNA after FTO knockdown, overexpression, or FTO inhibitor MA2 treatment. Aerosols), CDP-100-15 Chlorine Color Change Leak Detecting Paint (12oz Aerosol), ABCD-311-18 Acid Base PiNPOiNT Developer (12oz Aerosol), ABCD-311-15 Acid Base PiNPOiNT Developer (12oz Aerosol), ODP-340-15 Oil / Fuel Leak Detection Powder (Aerosol Can 12oz), Hydro-1-15 Color Change Developer (12oz Aerosol), WDP-217-18 Water Detecting Paint (Aerosol Cans 12), WDP-217-15 Water Leak Detecting Paint (aerosol), W-15A-M Water Leak Detection Monitors (4" x 4.25", pkg of 12). /Resources << /Type /Catalog FTO positively mediates the expression of miR181b3p downstream targets. /Type /Page You want to build livemaps for indoor use with multi-level settings? Wemap Pro includes a WYSIWYG tool to create, edit and import GeoJSON layers, immediately rendered in your livemaps.

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ErbB2driven breast cancer cell invasion depends on a complex signaling network activating myeloid zinc finger1dependent cathepsin B expression. 245 0 R null null null 246 0 R null null null 247 0 R null Screenshot: livebar opened / hidden in a mobile (vertical) screen, Screenshot: livebar opened on a wide screen. Briefly, 200ng of qualified RNA was added to the assay wells provided by the kit. /Keywords () Previous studies have demonstrated that ARL5B overexpression facilitated lysosome motility, resulting in lysosome dispersion and accumulation at the cell periphery [38, 39]. 145 0 R 146 0 R null null 236 0 R null null] /C2_0 45 0 R The closure of the gap was imaged at designated time intervals under 100 field, and the wound closure speed was evaluated using the ImageJ software. It is a customizable product that combines content and services in a geographical setting. Transfection was conducted using Lipofectamine 3000 (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA) according to the manufacturer's instructions. Aerosol)WDP-217 is an aerosol water leak detector/color change developer, which when sprayed on the points of suspected leaks (joints, welds, W-15A-M Water Leak Detection Monitors (4" x 4.25", pkg of 12)W-15A-M PinPoint Colormetric Paper for Water Leak Detection (4" x 4.25" adhesive back, pkg of 12 sheets to a carton). THW and PS designed and funded the study. NOTE: THIS PRODUCT CAN ONLY BE SHIPPED FEDEX GROUND SERVICES. >> A scatter plot of the relative correlation between FTO and ARL5B expression was performed using the R software. 250 0 R null null null 251 0 R null null null 252 0 R null 102 0 R 103 0 R 104 0 R 105 0 R 106 0 R 107 0 R 108 0 R 109 0 R 110 0 R 111 0 R /Parent 4 0 R /StructParents 1 Written informed consent was obtained from all patients. Either your web browser doesn't support Javascript or it is currently turned off. /Marked true In summary, these data indicate that FTO could accelerate the migration and invasion of HER2positive breast cancer cells via the FTO/miR181b3p/ARL5B signaling pathway (Figure10). [13] showed that FTO reduced the transcription of ankyrin repeat and SOCS box containing 2 (ASB2) and retinoic acid receptor alpha (RARA) by serving as an m6A demethylase, promoting proliferation and inhibiting apoptosis in acute myeloid leukemia. E Wound healing assay of SKBR3 and MDAMB453 cells pretreated with gradient concentrations of MA2 for 48 h. DMSO was used as solvent control. H Transwell invasion assay of SKBR3 and MDAMB453 cells transfected with FTO plasmid. Su etal. 4 Department of Basic Medicine, Zhongshan School of Medicine, Sun Yatsen University, Guangzhou Guangdong, 510080 P. R. China. Significant differences were calculated by twotailed Student's ttest or oneway analysis of variance (oneway ANOVA). My family immigrated to the USA in the late 60s. SHZ and KWZ provided supervision and suggestions for the implementation of the study. Due to the m6A demethylase function of FTO, we found that m6A was irregularly downregulated in HER2positive breast cancer tissues (Figure1C). A livemap contains several high-level, functional components that interact with each other. Further studies are urged to fully unveil the comprehensive role of FTO in breast cancer. Plasmid clones expressing FTO (oeFTO), ARL5B (oeARL5B), or empty vector (oeVec) were purchased from Sino Biological Inc. (Beijing, China). >> 130 0 R 131 0 R 132 0 R 133 0 R 134 0 R] By wound healing assay in SKBR3 cells and Transwell migration assay in MDAMB453 cells, cell migration was decreased upon FTO knockdown but accelerated upon FTO overexpression (Figure2C2D). Abbreviations: miRNC, negative control of miR181b3p mimics; miRNC2, negative control of miR181b3p inhibitors; ARL5B, ADP ribosylation factor like GTPase 5B; RAP2C, Rasrelated protein Rap2c; CLTA, clathrin light chain A; PCMTD1, proteinLisoaspartate (Daspartate) Omethyltransferase domain containing 1; RIMKLB, ribosomal modification protein rimK like family member B; SOWAHB, sosondowah ankyrin repeat domain family member B; other abbreviations as in Figure2. /Im2 40 0 R Ltd. is ISO 9001 Certified Company.Gas Leak Detection Sprays is an aerosol product that reveals the tiniest gas leaks in all sealed systems. 3 0 obj The preview is a media on the background (image thumbnail or icon) and a customizable subset of the pinpoints content in the livebar. Welcome to Iggy Garcia, The Naked Shaman Podcast, where amazing things happen. The livebar is highly customizable both in terms of User Interface (layout) and User Experience (behaviour). Screenshot - a livemap with third-party tiles in the background. stream /Type /Page This observation implied the intricate involvement of FTO in different breast cancer molecular subtypes, which is worthy of further exploration. The fully fledged editing tool allows you to define the parameters of your background style. Moreover, FTO promoted cell invasion and migration in vitro. Correlation analysis revealed that the expression levels of FTO and ARL5B were positively correlated (Figure7D), and the dual luciferase reporter assay confirmed that miR181b3p bound directly to the ARL5B 3UTR (Figure7E7F). Statistical analyses were performed using IBM SPSS Statistics version 22.0 (SPSS Inc., Chicago, IL, USA) and GraphPad Prism 7.0 (GraphPad Software Inc., La Jolla, CA, USA). A 10 L sterile pipette tip was used to scratch the cell layer and form a wound. Gas Leak Detection Sprays is an aerosol product that reveals the tiniest gas leaks in all sealed systems. 79 0 R null null 160 0 R null null null 161 0 R null null] /Contents [53 0 R 54 0 R 55 0 R] Received 2020 Jan 4; Revised 2020 Apr 22; Accepted 2020 Jun 27. /Manager (Publications) Correspondingly, FTO inhibition by siRNA or MA2 was found to impair cell invasion (Figure2F and2G), while the results were reversed when FTO was overexpressed (Figure2H). /TT3 34 0 R Conference Proceedings /MarkInfo << /S /Sect RNA sequencing (RNAseq) was conducted to decipher the downstream targets of FTO. This preview can be hidden depending on the amount of pinpoints in the visible (active) area of the map background. 208 0 R 209 0 R 210 0 R 211 0 R 212 0 R 213 0 R 214 0 R 215 0 R 216 0 R 217 0 R Our leak detection spray does not form harmful compounds with the following gases: For all compressed air, natural gas or liquid gas piping jobs, Detecting leakages on fittings, threaded joints, and connections, Detection of leaks in high-pressure pipes, valves, and compressed air cylinders. /P 5 0 R /MediaBox [0 0 595.32 842.04] In the present study, we discovered that FTO was highly expressed in HER2positive breast cancer cells and tissues. 1 Place Francis Ponge All these functions are offered in a feature rich and WYSIWYG interface, allowing a powerful and seamless way to create rich local information experiences tailored to your audiences needs. Abbreviations: ARL5B, ADP ribosylation factor like GTPase 5B; siARL5B, ARL5B siRNA; oeARL5B, ARL5B clone plasmid; other abbreviations as in Figure2, ARL5B plays an oncogenic role in HER2positive breast cancer cells. Pinpoints are geolocalized pieces of content present in the livemap. endobj Overexpressing ARL5B after MA2 treatment also rescued the inhibitory effect of MA2 (Figure8E and8F). Pan H, Gray R, Braybrooke J, Davies C, Taylor C, McGale P, etal. 2 /ParentTreeNextKey 7 /Rotate 0 Lysosomal trafficking, antigen presentation, and microbial killing are controlled by the Arflike GTPase Arl8b, Arl8 and SKIP act together to link lysosomes to kinesin1, The Arffamily protein, Arl8b, is involved in the spatial distribution of lysosomes, The small G protein Arl8 contributes to lysosomal function and longrange axonal transport in Drosophila. Zhou K, Sun P, Zhang Y, You X, Li P, Wang T. Estrogen stimulated migration and invasion of estrogen receptornegative breast cancer cells involves an ezrindependent crosstalk between G proteincoupled receptor 30 and estrogen receptor beta signaling, Overexpression of the fat mass and obesity associated gene (FTO) in breast cancer and its clinical implications. Furthermore, m6A quantification assay showed that inhibition of FTO increased m6A levels, while FTO overexpression decreased m6A abundance (Figure2I). /CropBox [0 0 595.32 842.04] /DropCap /Figure The authenticity of all cell lines was confirmed by short tandem repeat (STR) DNA profiling analysis. /F6 36 0 R /Parent 4 0 R Sold under the brand name of Spray Fill for more than 30 years. /TT3 35 0 R /Parent 4 0 R D:20100920202336 In the present study, miR181b3p inhibited the migration and invasion of HER2positive breast cancer cells by targeting ARL5B. Protein concentration was determined with the BCA protein assay kit (Thermo Fisher Scientific, Inc.). << All miR1813p mimics, inhibitors, and corresponding negative controls (miRNC, miRNC2) used for transfection were purchased from GenePharma Co., Ltd. (Shanghai, China). Steffan JJ, Williams BC, Welbourne T, Cardelli JA. << /TT1 33 0 R Li etal. The search engine is present natively on the livemap and provides the viewer with an additional ability to navigate the livemap and explore its content (pinpoints, events, lists). Knockdown and overexpression of FTO were carried out in SKBR3 and MDAMB453 cells, and corresponding changes in the expression of FTO were confirmed in both cell lines (Figure2A2B). Experiments were repeated independently at least three times. 1 Department of Physiology, Zhongshan School of Medicine, Sun Yatsen University, Guangzhou Guangdong, 510080 P. R. China. The optimal cutoff value of FTO (8.0) generated from the receiver operating characteristic (ROC) curve divided FTO expression into high or low. HGFinduced invasion by prostate tumor cells requires anterograde lysosome trafficking and activity of Na+H+ exchangers. To learn more about pinpoints, refer to entity description here. Dominissini D, MoshitchMoshkovitz S, Schwartz S, SalmonDivon M, Ungar L, Osenberg S, etal. /Font << Su R, Dong L, Li C, Nachtergaele S, Wunderlich M, Qing Y, etal. BC qRTPCR analysis of the expression of the six candidate molecules in SKBR3 and MDAMB453 cells with the transfection of miR181b3p mimics. The description and the details of the events are visible when the pinpoint is opened. /Kids [7 0 R 8 0 R 9 0 R 10 0 R 11 0 R 12 0 R 13 0 R] Rafn B, Nielsen CF, Andersen SH, Szyniarowski P, CorcelleTermeau E, Valo E, etal. << 14 0 obj /StructParents 3 C Relative m6A content of total RNA in paired HER2positive breast cancer tissues and adjacent nontumor tissues (n=12). 168 0 R 169 0 R 170 0 R 171 0 R 172 0 R 173 0 R 174 0 R 175 0 R 176 0 R 177 0 R 149 0 R 150 0 R 151 0 R 152 0 R null null 244 0 R null null null /Count 7 The contribution of FTO to the risk of various cancers shown in epidemiological studies [12], along with its aberrant expression in breast cancer, indicates the possible involvement of FTO in mediating the pathological development of breast cancer. Wound healing assay and Transwell assay were performed to determine the migration and invasion abilities of SKBR3 and MDA-MB453 cells with either knockdown or overexpression of FTO. We selected miR181b3p, the miRNA with the greatest change, to test the relationship between FTO and miRNAs in HER2positive breast cancer. Data are presented as the mean standard deviation. These results may imply the direct interaction between FTO and miR181b3p. /C2_0 31 0 R RNA sequencing (RNA-seq) was conducted to decipher the downstream targets of FTO. /ColorSpace << Tinghuai Wang, Email: nc.ude.usys.liam@htgnaw. 142 0 R 143 0 R 144 0 R 145 0 R 146 0 R 147 0 R 148 0 R 149 0 R 150 0 R 151 0 R Contact us. [23] searching for the possible consensus sequence motifs for m6A modification in m6A immunoprecipitated miRNAs, we succeeded in mapping the miR181b3p sequence to all the most discriminating motifs, and the mapped sequence was enriched in the center section. /TT0 32 0 R Abbreviations: FTO, fat mass and obesityassociated; HER2, human epidermal growth factor receptor 2; m6A, N6methyladenosine; HE, hematoxylin and eosin; IHC, immunohistochemistry; DFS, diseasefree survival; OS, overall survival; RFS, recurrencefree survival. /CS0 [/ICCBased 30 0 R] Wemap is a proud member of the OpenStreetMap community and our mapping data is synchronized with the OpenStreetMap dataset: this enables you to create your own style by choosing among numerous features and options including, among others: You have more specific needs? For the Transwell invasion assay, briefly, 5 1041 105 cells in 200 L serumfree DMEM were reseeded into the top of the insert of a Boyden chamber (Corning Inc., Corning, NY, USA) with 300g/mL Matrigel (BD Bioscience, San Jose, CA, USA), while 600 L medium with 10% FBS was loaded into the well below. Mechanistically, in SKBR3 and MDAMB453 cells, FTO accelerated cell migration and invasion by targeting ARL5B via miR181b3p downregulation. Schiefermeier N, Scheffler JM, de Araujo ME, Stasyk T, Yordanov T, Ebner HL, etal. >> /Font << The illumination of its regulatory mechanism will be helpful to clarify the clinical significance of miR181b3p in different circumstances [34]. background selection, within a catalogue of styles that are tailored for print (colorimetric adjustments, reduction of the geographic feature set to enhance readability on a printed material, etc. Abbreviations are the same as in Figure2, miR181b3p inhibits the protumor effects of FTO in HER2positive breast cancer cells. To explore potential targets of FTO, we performed RNAseq on FTOknockdown SKBR3 cells. Overall, ARL5B was identified as the downstream target of interest. Moreover, FTO promoted cell invasion and migration in vitro. /TT2 33 0 R Within Wemap Pro the Wemap Platform can be explored through a search engine that can be used to inspire and re-use content (pinpoints, lists, livemaps). /C2_0 45 0 R /Chart /Figure The binding site of miR181b3p to the ARL5B 3untranslated region (3UTR) was predicted using the miRNA databases mentioned above. Chua BA, Van Der Werf I, Jamieson C, Signer RAJ. /Type /Page Further studies are needed to fully unfold the clinical value of FTO, miR181b3p, and ARL5B in breast cancer. The background can also include some additional interactive layers (polygons, etc.). MicroRNAs (miRNAs) are a family of short noncoding singlestranded RNAs containing 1922 nucleotides that promote the degradation or translation inhibition of mRNA by targeting specific mRNAs and forming RNAinduced silencing complexes [19]. /Font << Gas Leak Detection Sprays can be used all types of plant, irrespective of the type of gas used.Several features are: We import Gas Leak Detection Sprays from FAREN Chemical Industries, Italy. /Rotate 0 The shaded area denoted the complementary bases and corresponding mutated nucleobases. All components are highly customizable, both in terms of User Interface and of feature-set. In addition, we observed inhibition of cell migration with MA2 treatment, and the inhibition effect was maximal at 80mol/L (Figure2E). Huang Y, Yan J, Li Q, Li J, Gong S, Zhou H, etal. 450) was measured using a microplate reader (SpectraMax M5 Molecular Devices, LLC, Sunnyvale, CA, USA) to calculate the m6A levels. /Parent 4 0 R Easy to clean off. D Representative IHC images of HER2 and FTO expression in tumor and nontumor tissues. /Author (European Space Agency) The manual / bulk creation of pinpoints from Wemap Pro offers a wide range of options to create rich content to be consumed from livemaps: media-rich pinpoints, events, html formatted descriptions, tags, synchronisation with social media accounts, and much more. or via an external application (app) that is synchronized with the Wemap platform. Therefore, 80mol/L MA2 was used for subsequent studies. Membranes were incubated with primary antibodies at 4C overnight and then secondary antibodies at room temperature for 2 h. After washing, signals were detected using a ChemiDoc imaging system (BioRad Laboratories, Inc.). Afterwards, the Venn diagram online tool (http://bioinformatics.psb.ugent.be/webtools/Venn/) was utilized to narrow down the scope into six target molecules. /StructParents 4 /Length 10 Ragvin A, Moro E, Fredman D, Navratilova P, Drivenes, Engstrm PG, etal. Lambert AW, Pattabiraman DR, Weinberg RA. For more details see livemap options and styling module. /Type /Page Colormetric detectors are rapid leak ABCD-311-18 Acid Base PiNPOiNT Colormetric Developer (12oz Aerosol [12 cans]) << Mechanistically, RNA-seq and further confirmation studies suggested that FTO up-regulated ARL5B by inhibiting miR-181b-3p. /Footnote /Note RNAseq analysis revealed that 19 miRNAs were significantly downregulated, while 10 miRNAs were significantly upregulated (Figure3A) the top six upregulated or downregulated miRNAs are listed in Figure3B. endstream >> >> On Wemap Pro a user will access its profile page to define or edit its personal information but also tailor her access rights, depending on the chosen Wemap subscription. Pinpoint utilizes only the most advanced electronic leak detection devices, including infrared thermal imaging, combined with proprietary techniques to pinpoint the exact location of your leak. In the present study, we confirmed that FTO could reduce the expression of miR181b3p, which inhibited the protumor effects of FTO. The results showed that FTO reinforced ARL5B expression, while miR181b3p inhibited ARL5B expression (Figure7A7C). /CS0 [/ICCBased 30 0 R] For more information, please visit: /TT2 52 0 R DE qRTPCR analysis of the expression of the six candidate molecules in SKBR3 and MDAMB453 cells with the transfection of miR181b3p inhibitors. /PageLayout /SinglePage Screenshot - Wemap Pro - explore the platform. Ltd. is ISO 9001 Certified Company. /ColorSpace << P value<0.05 was considered statistically significant. After 48 h of cotransfection, luciferase reporter assays were performed using the Dual Luciferase Reporter Assay System (Promega Corporation, Madison, WI, USA). As previously described [26], cultured cells were seeded into 24well plates at a density of 2 1054 105 cells per well. Cancer cells subcultured for less than 56 times were used in all experiments. From the water meter to anywhere in your home or building, including outside rain or moisture infiltration. Berulava T, Rahmann S, Rademacher K, KleinHitpass L, Horsthemke B. Ye S, Xu Y, Li J, Zheng S, Sun P, Wang T. Prognostic role of GPER/Ezrin in triple negative breast cancer is associated with menopausal status. stream uuid:e0c7ab79-8469-4935-9ee1-ca369aba8c50 We further verified that ARL5B also displayed carcinogenic activity in breast cancer cells.

Conclusion

Our work demonstrated the carcinogenic activity of FTO in promoting the invasion and migration of breast cancer cells via the FTO/miR-181b-3p/ARL5B signaling pathway. >L]"ZIa+~lEc' ${'8PBLO{. [44] detected that decreased lysosomal outward trafficking resulted in inhibited cancer cell invasion. /TT2 34 0 R AB The transfection efficiency of FTO siRNA or overexpression plasmid in SKBR3 and MDAMB453 cells confirmed by qRTPCR (A) and Western blotting (B). endobj stream << In the current study, we provide clinical and in vitro evidence that FTO, which is highly expressed in HER2positive breast cancer, could promote cell migration and invasion through the FTO/miR181b3p/ARL5B signaling pathway. /Type /Page /F8 38 0 R 1 [null null 162 0 R null null null 163 0 R null null null qRTPCR analysis confirmed that FTO inhibition using siRNA or MA2 resulted in significantly increased levels of miR181b3p, while FTO overexpression reduced miR181b3p expression (Figure4). The RTCGA. N6methyladenosine (m6A) modification, with methylated adenosine at the N6 position, is the most abundant internal modification of eukaryotic mRNA and noncoding RNA [7]. /StructParents 2 152 0 R 153 0 R 154 0 R 155 0 R] >> Moreover, based on the study by Berulava etal. >> /C2_0 31 0 R However, cell proliferation was not distinctly changed after FTO alteration (Supplementary Figure S1B). Screenshot - Airport livemap with multilevel management. endobj /TT1 52 0 R ADP-319-05 GAL PINPOINT COLORMETRIC- AMMONIA. Garg S, Sharma M, Ung C, Tuli A, Barral DC, Hava DL, etal. /Resources << /Type /Pages /Font << /TT0 32 0 R endobj /Rotate 0 /TextBox /Art 6 [null null 237 0 R null null null 238 0 R null null null /Outline /Span >> If youre curious about my background and how I came to do what I do, you can visit my about page. qRT-PCR, luciferase reporter assay, and Western blotting were employed to confirm the existence of the FTO/miR-181b-3p/ARL5B axis. /MediaBox [0 0 595.32 842.04] /Rotate 0 qRTPCR was performed using the SYBRGreen Master mix (Takara) in a CFX96 touch system (BioRad Laboratories, Inc., Hercules, CA, USA). Methyltransferase like 14 (METTL14) facilitates DiGeorge syndrome critical region 8 (DGCR8) recognition by adding m6A modification to primiR126, subsequently leading to the improvement of hepatic carcinoma [21]. Screenshot - Wemap Pro - livemaps management, Screenshot - Wemap Pro - livemap details management. The miR181 family has been clarified to be critical for cell cycle progression and differentiation [31]. All reads were mapped to human genome 19 using BWA tools and annotated by miRBase version 21 (https://www.mirbase.org) using miRDeep2 (https://www.mdc-berlin.de/8551903/en/). [27] discovered that FTO expression was significantly associated with clinicopathological characteristics of HER2positive breast cancer patients, which aligns with our current results. * P<0.05, ** P<0.01, *** P<0.001. /Endnote /Note Topology of the human and mouse m6A RNA methylomes revealed by m6Aseq. The followup deadline was September 20, 2019. If user has the access rights, the print module is available directly from Wemap Pro. 27 0 R 28 0 R 29 0 R] A Relative expression of FTO protein in breast cancer cell lines compared with the immortalized mammary epithelial cell line MCF10A. >> /C2_0 31 0 R The content on the livebar is always synchronized with the content displayed above the background to provide a consistent browsing experience. Screenshot: Livemap - web - mobile screen, Screenshot: livemap - web - desktop view - simplified chinese, Screenshot: livemap - mobile view - with itinerary, Screenshot: livemap - public screen (muse du Louvre). /Font << Experiments were terminated after scratch for 24 h. GH Transwell invasion assay in ARL5Bsilenced or overexpressing SKBR3 and MDAMB453 cells. endobj 16 0 obj B The top six upregulated or downregulated miRNAs. * P<0.05, ** P<0.01, *** P<0.001. In our present study, we failed to pinpoint the exact m6A modification site in miR181b3p by MeRIPseq. A methylated RNA immunoprecipitation sequencing (MeRIPseq) study showed that the steadystate levels of several miRNAs were affected by the knockdown of FTO and that a significant fraction of miRNAs contained m6A sites [23]. 30, Ground Floor, Ashoka Shopping Center LT Marg Mumbai - 400001, Maharashtra, India, No. The guided outward movement of lysosomes is considered to be a critical determinant of lysosomal function [42, 43]. The background component consists of a map made of tiles (vector or raster), hosted on Wemap servers and customizable (language, graphic styling, indoor mapping, etc.). 188 0 R 189 0 R 190 0 R 191 0 R 192 0 R 193 0 R 194 0 R 195 0 R 196 0 R 197 0 R 2 0 obj >> ARL5B is the target of miR181b3p and positively regulated by FTO. Experiments were terminated after seeding for 24 h. I Relative m6A content of total RNA after FTO knockdown, overexpression, or FTO inhibitor MA2 treatment. Aerosols), CDP-100-15 Chlorine Color Change Leak Detecting Paint (12oz Aerosol), ABCD-311-18 Acid Base PiNPOiNT Developer (12oz Aerosol), ABCD-311-15 Acid Base PiNPOiNT Developer (12oz Aerosol), ODP-340-15 Oil / Fuel Leak Detection Powder (Aerosol Can 12oz), Hydro-1-15 Color Change Developer (12oz Aerosol), WDP-217-18 Water Detecting Paint (Aerosol Cans 12), WDP-217-15 Water Leak Detecting Paint (aerosol), W-15A-M Water Leak Detection Monitors (4" x 4.25", pkg of 12). /Resources << /Type /Catalog FTO positively mediates the expression of miR181b3p downstream targets. /Type /Page You want to build livemaps for indoor use with multi-level settings? Wemap Pro includes a WYSIWYG tool to create, edit and import GeoJSON layers, immediately rendered in your livemaps. 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