MYC overexpression reduces myeloid differentiation of Asxl1- and CSF3R-mutant cells. (A) Heat maps representing the normalized, row-scaled counts of the top 50 most significantly differentially expressed genes (Padj < .05) between the control and double-mutant cells from the gene subset, Hallmark MYC targets. (B) Quantitative PCR (qPCR) for Myc before and after estrogen withdrawal (mean ± SEM; n = 3 per group). Significance was evaluated with a 2-way ANOVA with Sidak multiple comparison test. (C) qPCR for Myc expression in transduced double-mutant cells with a Tet-on MYC plasmid (mean ± SEM; n = 3 per group). Significance was evaluated with a 2-way ANOVA with Sidak multiple comparison test. (D) Representative flow cytometry plots of CD11b and GR1 at 48 hours post estrogen withdrawal of the double-mutant cells with a Tet-on MYC plasmid. These cells were treated with or without doxycycline 24 hours before estrogen withdrawal. Quantification of CSF3R-mCherry/Asxl1-GFP–positive cells reveals significant repression of CD11b⁺/GR1⁺ (mean ± SEM; n = 3 per group). Significance was evaluated with a 2-way ANOVA with Sidak multiple comparison test. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001.