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. Author manuscript; available in PMC: 2022 Mar 18.
Published in final edited form as: Mol Cell. 2021 Aug 5;81(17):3604–3622.e10. doi: 10.1016/j.molcel.2021.07.018

Figure 4. ZMYND8 regulates IRF8 and MYC transcription to sustain AML proliferation.

Figure 4.

(A) Schematic depicting establishment of an inducible ZMYND8 degradation system in MOLM-13 cells.

(B) Competition-based proliferation assay of MOLM-13-dZD8 versus parental cells.

(C) RNA-seq analysis of gene expression changes in MOLM-13-dZD8 cells treated with either DMSO or 500 nM dTAG-13 for 4 h (n = 2).

(D) Time-course reverse transcriptase quantitative PCR (RT-qPCR) analysis of mRNA expression in MOLM-13-dZD8 cells treated with 500 nM dTAG-47. Relative mRNA levels were normalized to GAPDH levels.

(E) RNA-seq analysis of gene expression changes 5 days after transduction of sgNeg or sgZMYND8. Myeloid-differentiation-associated genes are labeled in blue.

(F) Immunoblotting of ZMYND8, MYC, IRF8, or GAPDH in whole-cell lysates.

(G) GSEA of RNA-seq data presented in Figure 4A. Myc_Targets_Up_Schuhmacher (Schuhmacher et al., 2001) and IRF8_Targets_Up or Myeloid_development_Up (Brown et al., 2006) signatures were used.

(H) Competition-based proliferation assays performed in MOLM-13 cells expressing EV, MYC, IRF8, or MYC+IRF8 and transduced with indicated sgRNAs. EV, empty vector.

Data points in line graphs represent the average of three independent biological replicates (n = 3). Error bars represent mean ± SEM. See also Figure S4 and Table S1.