Fig. 4. CARM1 negatively regulates MLL3 interaction with ASXL2/BAP1 and impairs its chromatin binding and gene expression.

(A) Plasmids expressing the WT and mutant LOCAP domain were transfected in HEK293T cells for 24 hours. The protein-protein interaction between MLL3 and the LOCAP domain was determined by an IP experiment. (B) Endogenous IPs with MLL3 antibody from CARM1-WT and CARM1-KO cells were subjected to Western blotting with BAP1 and ASXL2 antibody; n = 2. (C) The log₂FC heatmap showing MLL3 occupancy in BAP1-KO and CARM1-KO cells. (D) The average plot compares MLL3 occupancy between WT cells, BAP1-KO cells, and CARM1-KO cells. (E) The ChIP-seq track example shows an increased in MLL3 occupancy at enhancers due to CARM1 depletion (CARM1-KO). The RNA-seq heatmap (F) and track example (G) shows the gene expression change between BAP1-, MLL3-, and CARM1-depleted cells.