Project description:Macrophages are cells belongs to innate immune system, which response to pathogen by the production of inflammatory proteins those that are effective in both combating pathogen and wound healing. Using small molecule inhibitors it has been shown that many of the inflammatory response genes were under control of BET proteins. We used LysM-Cre to conditionally delete floxed BRD4 in resident peritoneal macrophages and monitored the effect on inflammatory gene expression
Project description:We are reporting here genome wide distribution of BRD4 in bone marrow derived macrophages at steady state level and in reponse to LPS stimulation. In both condition, BRD4 distribution was found to be widespread in intragenic as well as intergenic regions with enrichment specifically near TSS. Characteristically BRD4 signal were highly enriched on enhancers of some active genes and hence were classified as BRD4 rich super-enhancers (>12kb long stretches). Super-enhancers are formed, in some instances by redistribution of BRD4 on LPS stimulated genes. BRD4 binding well correlated PolII binding at the chromatin acetylated at H3K27, H3K9 and H4tetraK. Interestingly many LPS induced BRD4 independent genes, despite BRD4 loss in the KO, maintained PolII binding at the chromatin enriched with acetylated lysines at H3K27, H3K9 and H4tetraK. Additionally among some BRD4 independent LPS inducible genes we detected stronger p65 enrichment in the KO. In contrast, P65 binding was not detected in BRD4 dependent genes. These observations provided evidence of plasticity of epigenetic changes in retaining inflammatory reponses in macrophages.
Project description:Macrophages are cells belongs to innate immune system, which response to pathogen by the production of inflammatory proteins those that are effective in both combating pathogen and wound healing. Using small molecule inhibitors it has been shown that many of the inflammatory response genes were under control of BET proteins. We used LysM-Cre to conditionally delete floxed BRD4 in bone marrow derived macrophages and monitored it's effect on inflammatory gene expression
Project description:Genome-wide BRD4, PolII and p65 occupancy as well as chromatin modifications in bone marrow derived macrophages from BRD4 conditional KO mice
Project description:Next Generation Sequencing (NGS) of expression profile from unstimulated and LPS stimulated bone marrow derived macrophages from control and BRD4 conditional KO mice
Project description:To investigate the role of BRD4 in normal hematopoiesis, Brd4 conditional KO mouse model was generated. We purified the Lin-cKit+ cells from bone marrow of WT and Brd4 KO mice and performed the CUT&RUN to study the histone modification and Brd4 occupancy around genome in HSC/HPCs population.
Project description:To investigate the role of BRD4 in normal hematopoiesis, Brd4 conditional KO mouse model was generated. We purified the Lin-cKit+ cells from bone marrow of WT and Brd4 KO mice and performed the ATACseq to study the chromatin accessibility in each cell population.