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. Author manuscript; available in PMC: 2022 May 1.
Published in final edited form as: Cancer Discov. 2021 May 12;11(11):2868–2883. doi: 10.1158/2159-8290.CD-20-1793

Figure 6.

Figure 6.

MYB interference downregulates EVI1 but not GATA2.

A. Western blot for MYB, EVI1 and GATA2 in MUTZ3-EVI1-GFP upon sgRNA-mediated MYB knockout (MYB.30) at indicated days after induction of Cas9. Actin was used as loading control.

B. Western blot for MYB, EVI1 and GATA2 in untreated cells (−) or cells treated for two days with 20 μM of TG3 or MYBMIM (MM). Actin was used as loading control.

C. Colony forming units (CFU) of MUTZ3 cells cultured without peptide or treated with 20 μM TG3 or MYBMIM for two days and subsequently plated in methylcellulose. Error bars show standard deviation across three plates. P-values were calculated using a one-way ANOVA test.

D. Flow cytometric analysis of MUTZ3 cells stained with CD34 and CD15. Cells studied by flow cytometry were either untreated or treated with 20 μM TG3 or MYBMIM for two days and subsequently grown for nine days in methylcellulose.

E. Colony forming units (CFU) of MUTZ3 cells with pMY-FLAG-Evi1-IRES-GFP (Evi1) or empty vector (EV) cultured without peptide or treated with 20 μM MYBMIM for two days and subsequently plated in methylcellulose. Error bars show standard deviation across three plates. P-values were calculated using a one-way ANOVA test.

F. Flow cytometric analysis of MUTZ3 cells with Evi1 or EV, stained with CD34 and CD15. Cells studied by flow cytometry were either untreated or treated with 20 μM MYBMIM for two days and subsequently grown for eight days in methylcellulose.

G. p300 and MYB ChIP-seq profiles of the 18 kb region in MUTZ3 cells treated with either 20 μM TG3 or MYBMIM for 48 h.

H. Cell-viability test of inv(3)/t(3;3) AML primary cells determined by CellTiter-Glo three days after culturing the cells in a 96-well plate with 20 μM TG3 or MYBMIM. Error bars show standard deviation across four biological replicates. P-values were calculated using a one-way ANOVA test.

I. Western blot for MYB, EVI1 and GATA2 in untreated AML cells or in AML cells treated with 20 μM TG3 or MYBMIM for 48h. Actin was used as loading control.

J. Western blot for MYB, EVI1 and GATA2 in cultured CD34+ cells untreated or treated with 20 μM TG3 or MYBMIM for 48h. Actin was used as loading control.