Project description:Purpose: The goals of this study are to compare transcriptome profiling (RNA-seq) resulting from a Mesp1Cre driven ablation of Hira in the heart at E11.5 and E12.5. Methods: RNA extraction was done in triplicate from Mesp1Cre;Hira-/fl and Mesp1Cre;Hira+/fl embryonic hearts at E11.5 and E12.5 using the QIAGEN RNeasy mini kit. RNAseq was processed by the ICH genomics facility, reads were aligned and normalised using BOWTIE and DEseq R package. Strand NGS 2.5 software was used to re-analyse the aligned files (.bam). By applying the Mann Whitney unpaired test, Benjamini Hochberg False discovery rate (FDR) and filtering the genes using adjusted p-value ≤ 0.05 and absolute fold change ≥ 1.5, 95 % of the results were identical to the DEseq package used by the UCL Genomics facility. Results: We identified 360 coding transcripts changed in the mutant hearts (Mann Whitney unpaired test, Benjamini Hochberg FDR, p ≤ 0.05, FC ≥ 1.5) with no trend towards up- or down-regulation of global transcription (48.8% down vs 51.2% up) at E12.5. Conclusions: This work analyses the role of HIRA in mouse cardiac development. It was found that Tnni2 is the most upregulated gene in the absence of Hira. qRT-PCR validation of 25 targets. Little (<5%) or no variation of fold change between RNAseq and qRT-PCR data were observed.
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from Mus musculus tissues (Heart, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:ATAC-seq and RNA-seq were used to compare the chromatin states and corresponding transcriptomes of developing HSPC. We found that the chromatin states and transcriptomes change markedly when HSPCs migrate from PL to FL and from FL to BM. Then we compared chromatin among E12.5 PL, E12.5 FL, and E16.5 FL, the proportion of fragments in the nucleosome-free region significantly increases in E12.5 and E16.5 FL-HSPCs compared with E12.5 PL-HSPCs; the motifs of ETS and Runt are enriched at the opeing or closing peaks from E12.5 PL to E12.5 FL and E12.5 FL to E16.5 FL, and the expression of these factors is either up-regulated or down-regulated; the motifs of bZIP and Zf are enriched at the closing and opening peaks from E12.5 PL to E12.5 FL and E12.5 FL to E16.5 FL respectively, In line, the expression of these factors is down-regulated or up-regulated.
