Project description:Purpose: we used next generation sequencing to analyze gene expression profiles of MEF treated with PBS (control), TNF-ɑ + Cycloheximide (TC) or TNF-ɑ + Cycloheximide+ z-VAD-fmk (TCZ). The goals of this study are to compare the different gene expression profiles between MEF treated with TC compared to PBS treatment and MEF treated with TCZ compared to PBS treatment. Methods: In MEF, the combination of TNF-α and cycloheximide (TC) induces apoptosis, whereas the addition of z-VAD-fmk (TCZ) to this regimen promotes necroptosis, using High-seq 2000 Illumina sequencing platform. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: By comparing the RPKM values between the two groups, a total 492 DEGs (342 upregulated and 150 downregulated) were identified in MEF treated with TC compared to PBS treatment, and a total 424 DEGs (242 upregulated and 182 downregulated) were identified in MEF treated with TCZ compared to PBS treatment. Next, we compared all identified DEGs in MEF between TC and TCZ treatment. We found that 252 DEGs overlapped between TC and TCZ treatment, whereas 240 DEGs were only present in TC treatment and 172 DEGs were only present in TCZ treatment. Conclusions: we firstly identified 492 DEGs in apoptotic MEF, and as well as 424 DEGs in necroptotic MEF. And we performed the enriched molecular pathway, molecules, transcription factors and signaling networks by IPA analysis. We revealed that apoptosis and necroptosis may share certain common canonical pathways and transcription factors.
Project description:This experiment is to compare derived iPSCs with original MEFs and real mES to test the overall quality of iPSCs and whether they are similar to mES on global mRNA expressions
Project description:PRDM proteins belong to the SET domain protein family, which is involved in the regulation of gene expression. Although few PRDM members possess histone methyltransferase activity, the molecular mechanisms by which the other members exert transcriptional regulation remain to be delineated. In this study, we find that Prdm5 is highly expressed in mouse embryonic stem (mES) cells and exploit this cellular system to characterize molecular functions of Prdm5. By combining proteomics and next-generation sequencing technologies, we identify Prdm5 interaction partners and genomic occupancy. We demonstrate that although Prdm5 is dispensable for mES cell maintenance, it directly targets genomic regions involved in early embryonic development and affects the expression of a subset of developmental regulators during cell differentiation. Importantly, Prdm5 interacts with Ctcf, cohesin, and TFIIIC and cooccupies genomic loci. In summary, our data indicate how Prdm5 modulates transcription by interacting with factors involved in genome organization in mouse embryonic stem cells. ChIP-Sequencing of fragments enriched by immunoprecipitation of Prdm5 in wild type mouse embryo fibroblasts (MEF) in duplicate. Background noise details obtained by ChIP-Sequencing of Prdm5 lacz/lacz MEF, in duplicate.
Project description:This SuperSeries is composed of the following subset Series: GSE38158: mes-2, mes-4 or mes-2; mes-4 mutants vs. wild type GSE38159: Strome MES-4, H3K36me3 and H3K27me3 in mes-4 RNAi EEMB GSE38180: Strome Mes-4, H3K36me3 and H3K27me3 in N2 EEMB Refer to individual Series
Project description:Trascriptional profiling of C. elegans adult germ lines comparing mes-2(bn11)unc-4(e120) mutants and unc-4(e120) (wild type), mes-4(bn85) mutants and N2 (wild type), mes-2(bn11)unc-4(e120); mes-4(bn85) and unc-4(e120) (wild type) at 20 degrees. One-condition experiment, mutant vs. WT. Biological replicates 4 mutant, 4 wildtype harvested indepedently. One mutant replicate and one WT replicate per array.
Project description:The microarray analysis was used for comparing the gene expression profiles between MEF, MEF treated with n-Butylenephthalide (BP) 10 and 40ug/ml.
