Project description:Hela test samples prepared by FASP digestion were run on QE and QE-HF mass spectrometry as quality controls. We tried to compare the protein and peptide identification results between the two machines.
Project description:PPARγ is a master transcriptional regulator of adipogenesis. Hence, the identification of PPARγ coactivators should help reveal mechanisms controlling gene expression in adipose tissue development and physiology. We show that the non-coding RNA Steroid receptor RNA Activator, SRA, associates with PPARγ and coactivates PPARγ-dependent reporter gene expression. Overexpression of SRA in ST2 adipocyte precursor cells promotes their differentiation into adipocytes. Conversely, knockdown of endogenous SRA inhibits 3T3-L1 preadipocyte differentiation. Microarray analysis reveals hundreds of SRA-responsive genes in adipocytes, including genes in cell cycle, insulin and TNFα signaling pathways. Some functions of SRA may involve mechanisms other than coactivation of PPARγ. SRA increases insulin-stimulated glucose uptake in adipocytes. SRA promotes S-phase entry during mitotic clonal expansion, decreases expression of cyclin-dependent kinase inhibiters p21Cip1 and p27Kip1, and increases phosphorylation of Cdk1/Cdc2. SRA also inhibits the TNFα-induced phosphorylation of c-Jun NH2-terminal kinase. In conclusion, SRA enhances adipogenesis and adipocyte function through multiple pathways.
Project description:We performed Chromatin Isolation by RNA Purification (ChIRP) of SRA and ChIP of p68 following by high-throughput sequencing in NTERA2 cell line. We find that SRA localizes with p68 genome-wide at genes whose function is involved in embryonic development.
Project description:Using the TempO-Seq rat S1500+ platform we performed gene expression analysis of using 63 purified RNA samples from the livers of rats exposed to controls or chemicals that fall into one of five modes of action (MOAs): constitutive androstane receptor/pregnane X receptor (CAR/PXR) activation, aryl hydrocarbon receptor (AhR) activation, peroxisome proliferator-activated receptor-alpha (PPARA) activation, cytotoxicity or DNA damage. The TempO-Seq data generated was used to compare to gene expression data acquired from the same samples run on Affymetrix microarays (GEO: GSE47875) and Illumina RNA-Seq (GEO: GSE55347, SRA:SRP039021).
