Figure 6.

ATAC maintains self-propagation and SAGA blocks differentiation of MOLM13 AML cells. (A) Quantification of flow cytometry analysis of cell cycle in MOLM-13 cells transduced with KAT2Ash, SAGA-specific SUPT20Hsh and TADA2Bsh, and ATAC-specific ZZZ3sh and TADA2Ash. Corresponding representative plots in supplemental Figure 5C. Mean ± SEM of 3 independent experiments. Two-tailed Student t test for significance *P < .05, **P < .01. (B) Representative flow cytometry plots of divisional tracking of MOLM-13 cells transduced with CTRLsh, SUPT20Hsh, and ZZZ3sh and loaded with the Tag-IT violet dye (Biolegend) after 3 days of culture. Regions 0, 1, and 2 represent the number of cell divisions relative to initial loading control (also see supplemental Figure 5D-E). (C) Quantification of results in panel B representing the distribution of MOLM-13 cells transduced with CTRLsh, SUPT20Hsh, and ZZZ3sh undergone 0-2 cell divisions after 3 days in culture. Mean ± SEM of 3 independent experiments. Two-tailed Student t test for significance *P < .05. (D) Colony replating of MOLM-13 cells transduced with SUPT20Hsh1, SUPT20Hsh2 (SAGA), TADA2Ash1, and TADA2Ash2 (ATAC) as a measure of in vitro self-renewal. Mean ± SEM of 2-3 independent experiments. Two-tailed Student t test for significance *P < .05. (E) Quantification of blast-like and differentiated cells in MOLM-13 cultures transduced with CTRLsh, KAT2Ash, SUPT20Hsh, and ZZZ3sh. Scoring of 3 randomly selected fields of >100 cells; Two-tailed Student t test for significance; *P < .05, **P < .01. (F) Representative photographs of MOLM-13 cytospins. White arrow heads denote blast-like cells; black arrow heads denote differentiated cells. Bar represents 50 μm. (G) Quantification of Annexin V⁺ apoptotic cells in MOLM13 cultures analyzed by flow cytometry (representative plots in supplemental Figure 4F). Mean ± SEM of n > 3 independent experiments. Two-tailed Student t test for significance.