Project description:Rbfox2 is required for maintaining hepatic cholesterol levels under high fructose. Here we carry out an enhanced iCLIP (eiCLIP) protocol in order to determine the targets of Rbfox2 in primary mouse hepatocytes.
Project description:We generated a global analysis of Rbfox2 splicing regulation combined with a highly specific, single nucleotide-resolution Rbfox2 RNA binding map. We found that Rbfox2 regulates the splicing and expression of many previously unknown targets, and particularly a number of RNA binding proteins (RBPs), by modulating alternative splicing coupled-NMD. Based on our observations of RBP-Rbfox2 co-regulation with a polarity predicted by Rbfox2 binding, we propose a model whereby Rbfox2 tunes autoregulatory splicing events to control RBP expression levels and in turn alter their respective splicing networks. iCLIP for epitope-tagged Rbfox2 and control untagged Rbfox2; RNAseq of control and Rbfox2 knockdown in mouse embryonic stem cells
Project description:Mouse primary hepatocytes (MPH) prepared from C57Bl6 male adult mice were challenged with the FXR agonist GW4064 and used to prepare RNA which was then processed for analysis using MoGene-2_0-st Affymetrix microarrays according to standard procedures.
Project description:Rbfox proteins regulate alternative splicing, mRNA stability and translation. These proteins are involved in neurogenesis and have been associated with various neurological conditions. We generated expression profile in adult and developing mouse retinas which lacks in Rbfox2 expression by RNA sequencing. The goals of this study is to identify the affected pathways in Rbfox2 KO mouse retina as well as potential splicing targets of Rbfox2.
Project description:Isolated mouse primary hepatocytes were cultured at glucose- and phenol-free DMEM medium, supplemented with 10mM pyruvate sodium and 10mM sodium lactate, and the cells were incubated for 1hr with vehice, recombinant mGP73 (64nM), or glucagon (3μM) ,cells were washed twice with cold PBS and scraped with cold RIPA lysis buffer supplemented with proteases and phosphatases inhibitors. Phosphoproteomics assay was performed by Shanghai luming biological technology co., LTD (Shanghai, China).