Study of MED1 acetylation in ER positive breast cancer cells [RNA-Seq IV]
Chromatin occupancy of MED1 and other Mediator components, as well as Pol II, were studied in MCF7 cells. MED1 acetylation affects chromatin occupancy of its related factors, which was analyzed with CUT&RUN-seq, ChIP-seq and sequential ChIP-seq assay (by pan-acetyl-lysine antibody) with MED1 KR mutant (HA tag) where acetylation residues were replaced. Combined with RNA seq analysis for gene expression in MCF7 cells with MED1 KO and KR/KQ MED1 ectopic expression, the role of MED1 acetylation in gene transcription control was investigated. Overall design: ChIP-seq and/or CUT&RUN-seq for MED1 and other Mediator compoments, as well as Pol II, were conducted in MCF7 cells. In addition to normal MCF7 cells, the cells with depletion of endogenous MED1 and ectopic expression of WT or KR mutant MED1 (HA tag) were also tested. Sequential ChIP-seq by FLAG (for MED1) antibody and pan-acetyl-lysine antibody were conducted. RNA seq (for mature mRNA) was conducted for MCF7 cells with MED1 KO (vs control), and 6KR/Q MED1 ectopic expression (Vs WT MED1 expression).