Project description:label the cells overexpressed Myc tagged METTL3 and Flag tagged WTAP with 4-SU, the RNA bound by METT3,WTAP can be got by Myc or Flag IP followed by RNA isolation by using the TRIzol (Invitrogen) reagent by following the company manual.the RNA was then used for library preparation using a TruSeq™ RNA Sample Prep Kit v2 (Illumina, San Diego, CA, USA) according to the manufacturer’s protocol.The libraries were sequenced using HiSeq2000 (Illumina) in single-read mode, creating reads with a length of 50 bp. Sequencing chemistry v2 (Illumina) was used and samples were multiplexed in two samples per lane. discovery of the binding motif of METTL3，WTAP in METTL3，WTAP overexpressed Human 293T cells

2014-01-16 | E-GEOD-50580 | biostudies-arrayexpress

The effects of WTAP on BAT function

Project description:Purpose: The goal of this study is to compare transcriptome profilings of interscapular brown adipose tissue (iBAT) from BAT-specific WTAP knockout and control mice Methods: iBATs were collected from WTAP BKO and f/f mice at 8 weeks old (n=3 for each group), and total RNA was isolated using TriPure. RNA-seq was performed by using Illumina NovaSeq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome (Ensemble_grcm38_p6) with TopHat (version 2.0.12), and the aligned reads were used to quantify mRNA expression by using HTSeq-count (version 0.6.1). Conclusion:Our study represents the first detailed analysis of iBAT transcriptomes from WTAP BKO and f/f mice, generated by RNA-seq technology. The RNA-seq analysis showed that 894 genes were down-regulated and 1410 genes were up-regulated in the iBAT of WTAP BKO mice. GO analysis showed that the down-regulated genes were primarily related to related to generation of precursor metabolites and energy, cellular respiration, energy derivation by oxidation of organic compounds, electron transport chain, nucleotide metabolic process and purine ribonucleotide metabolic process.

2022-10-19 | GSE202510 | GEO

The effects of WTAP on islet β-cell function

Project description:The effects of WTAP on islet β-cell function

| PRJNA889006 | ENA

The effects of WTAP on islet β-cell function

Project description:The goal of this study is to investigate how WTAP regulates islet β-cell function. Islets were isolated from pancreatic islets of Wtapflox/flox and Wtap-βKO mice at 7 weeks old. One islet sample was combined from three mice. Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany). Three independent biological replicates for each group were used for RNA-seq. RNA-seq was performed by deep sequencing using an Illumina Novaseq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome(GRCm38/mm10) with Hisat2 v2.0.5, and the aligned reads were used to quantify mRNA expression by using featureCounts v1.5.0-p3. Our study represents the first detailed analysis of islet transcriptomes from Wtapflox/flox and Wtap-βKO mice, generated by RNA-seq technology. The RNA-seq analysis showed that 3015 genes were downregulated and 2900 genes were upregulated in the pancreatic islets of Wtap-βKO mice.

2023-01-31 | GSE215156 | GEO

mRNA profiles of WTAP siRNA-treated HUVECs

Project description:To examine the role of WTAP in splicing regulation, we performed high-throughput mRNA sequencing (RNA-seq) on RNA isolated from control, WTAP or Virilizer siRNA-treated HUVECs, yielding 12 million uniquely mapped 75nt pair-end tags from each sample. MapSplice software was used for differential expression and differences in transcript splice junctions . mRNA profiles of control, WTAP or Virilizer siRNA-treated HUVECs were generated by deep sequencing using Illumina GAII.

2014-09-25 | E-GEOD-41180 | biostudies-arrayexpress

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