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. 2017 Apr 8;45(11):6388–6403. doi: 10.1093/nar/gkx234

Figure 5.

Figure 5.

MLL3/MLL4 are required for CBP/p300 binding on enhancers activated in adipogenesis. Mll3−/−Mll4f/f brown preadipocytes were infected with adenoviral GFP or Cre as described (5). Cells were collected at D2 of adipogenesis for ChIP-Seq of CBP and p300. Cells were collected at D0 and D2 of adipogenesis for western blot analysis of MLL4, CBP and p300. (A–C) MLL3/MLL4 facilitate CBP/p300 binding on active enhancers at D2. (A) Aggregated profiles. (B) Heat maps. (C) Deletion of Mll4 causes more severe impairment of CBP/p300 binding at MLL4+ than at MLL4− CBP/p300 sites on active enhancers. (D) Western blot analysis of MLL4, CBP and p300 at D0 and D2. RbBP5 serves as the loading control. (E) Genome browser view of enhancers (highlighted in gray) around the Pparg locus, where the deletion of Mll3/Mll4 resulted in decreases of CBP and p300 binding at D2. (F) MLL4 physically interacts with UTX and RbBP5 but not CBP/p300 during adipogenesis. Nuclear extracts of cells collected at D2 of adipogenesis were incubated with MLL4, CBP or p300 antibody. Immunoprecipitates were analyzed by western blot using antibodies indicated on the right.