MTCH2 knockdown increase histone acetylation, which is functionally important for the differentiation of AML cells and independent of BID. (A) Histones were isolated from control and MTCH2 knockdown clones of OCI-AML2, TEX, and 8227 cells. Levels of total and acetylated histones H3 and H4 were measured by immunoblotting. Representative immunoblots are shown. (B) NSE staining of OCI-AML2 cells transduced with shRNA targeting MTCH2 with or without cotreatment with C646 (5 µM) for 4 days. Quantitative analysis of NSE staining was performed with ImageJ software. (C-D) Proliferation curves of OCI-AML2 (C) and TEX cells (D) transduced with shRNA targeting MTCH2 shRNAs with or without C646 (5 µM). Levels of total and acetylated H3 histones were measured in OCI-AML2 (C) and TEX cells (D) after MTCH2 knockdown and cotreatment with 5 µM C646 for 4 days. (E) Venn diagram of the number of peaks and overlapping peaks detected by MACS2 in control, MTCH2 knockdown, and MTCH2 knockdown cells treated with C646. (F) Bar graphs of total numbers of peaks of H3K27ac in control, MTCH2 knockdown, and MTCH2 knockdown TEX cells treated with C646, detected by MACS2. (G) The bar plot represents the number of H3K27ac peaks in control, MTCH2 knockdown, and MTCH2 knockdown cells treated with C646 that overlap with ±6-kb regions centered at the transcription start site (TSS) of the 40 LSC⁻ genes that are upregulated in MTCH2 knockdown cells compared with control. (H) The average profiles and heat maps of the H3K27ac ChIP-seq signal in control, MTCH2 knockdown, and MTCH2 knockdown cells treated with C646 at ± 6kb of TSS of the 40 myeloid-like (LSC⁻) genes. The relative area under the curve (AUC) of TSS ± 6-kb regions is shown as bar graphs. (I) Histones were isolated from OCI-AML2 and TEX cells treated with or without Bid inhibitor, Bi-6C9, at Fas-inducing conditions (treated with CHX and CH-11). Levels of total and acetylated histones H3 and H4 were measured by immunoblotting. Representative immunoblots are shown.