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. 2022 Jan 7;6(1):165–180. doi: 10.1182/bloodadvances.2020002842

Figure 5.

Figure 5.

ATAC and SAGA differentially impact histone acetylation and gene expression in MOLM13 AML cells. (A) qRT-PCR validation of KAT2A, SUPT20H, TADA2B (SAGA-specific subunits), ZZZ3, and TADA2A (ATAC-specific subunits) knockdown in MOLM-13 cells. N ≥3 independent experiments, mean ± SEM of gene expression relative to CTRLsh, normalized to HPRT1 housekeeping gene. Two-tailed Student t test for significance *P < .05, **P < .01, ****P < .0001. (B) qRT-PCR analysis of ribosomal protein gene expression in MOLM-13 cells transduced with KAT2Ash, SAGA-specific SUPT20Hsh and TADA2Bsh, and ATAC-specific ZZZ3sh and TADA2Ash. N ≥3 independent experiments, mean ± SEM of gene expression relative to CTRLsh, normalized to HPRT1 housekeeping gene. Two-tailed Student t test for significance *P < .05, **P < .01, ***P < .001, ****P < .0001. (C) qRT-PCR analysis of self-renewal gene signature in MOLM-13 transduced with KAT2Ash, SAGA-specific SUPT20Hsh and TADA2Bsh, and ATAC-specific ZZZ3sh and TADA2Ash. N = 2 biological replicates, each run as 2 or 3 technical repeats; mean ± SEM of relative gene expression relative to CTRLsh, normalized to HPRT1 housekeeping gene. Two-tailed nested Student t test for significance *P < .05, ****P < .0001. (D) H3K9ac ChIP-qPCR analysis of ribosomal protein genes and self-renewal genes in MOLM-13 cells upon knockdown of SAGA and ATAC elements SUPT20H and ZZZ3, respectively. N ≥ 3 independent experiments. Mean ± SEM of enrichment relative to rabbit IgG, with normalization to control region in KRT5 locus with no significant H3K9ac enrichment. Two-tailed Student t test for significance *P < .05, **P < .01.