RUNX1-RUNX1T1 and GCN5 opposingly regulate CBFA2T3 transcription. Scatter plot showing fold changes of CBFA2T3 and GCN5 between primary and relapsed samples in both GSE66525 (A) and GSE83533 (B) patient cohorts. The red-colored dots in panel B denote 2 inv(16) patients. (C) Gene expression of CBFA2T3 and p21 in Kasumi-1, NOMO-1, and A549 cells treated with GCN5 inhibitor MB-3 vs dimethyl sulfoxide (DMSO). Results were analyzed from a public ERP003933 dataset.51 (D) RT-qPCR results showing the effects of CPTH2 and MB-3 treatment on CBFA2T3 and p21 expression in 2 primary AML patient samples. (E) ChIP-Seq intensities of RUNX1-RUNX1T1, HEB, E2A, GCN5, and acetyl-H3K9 at CBFA2T3 promoter sites in control (shControl) and RUNX1-RUNX1T1-depleted (shRUNX1-RUNX1T1) Kasumi-1 cells. (F) ChIP-qPCR quantification of the binding of the indicated proteins to −71 and −2021 CBFA2T3 regulatory loci in shControl and shRUNX1-RUNX1T1-treated Kasumi-1 cells. (G) RT-qPCR results of CBFA2T3 levels in control and RUNX1-RUNX1T1-knockdown Kasumi-1 cells with and without CPTH2 treatment. #P < .1, *P < .05, **P < .01; ***P < .001.