PRC1.1 is required for menin-inhibitor-induced antileukemia effects. (A) Scatter plots showing gene log2 FC for each gene in 2 biological replicates of CRISPR screen in MOLM-13 cells. (B) MAGeCK results showing ranked genes based on the RRA score of the day 14 VTP50469-treated group compared with the DMSO-treated group in MOLM-13 cells. (C) Western blot analysis showing the levels of the indicated proteins after individual knockout of PRC1.1 complex components BCOR, RYBP, and PCGF1 in OCI-AML2 cells. Two independent sgRNAs were used for targeting each gene. (D-F) Dose-response curves showing the viability of OCI-AML2 cells expressing the indicated sgRNAs, including PCGF1 (D), BCOR (E), and RYBP (F), after a 10-day treatment with the DMSO control or various doses of VTP50469. All cell viabilities were normalized to DMSO treatment. The calculated IC50 values are shown in boxplots. (G) Flow cytometry analysis of the percentages of CD11b+ cells in OCI-AML2 cells expressing sgLuc and sgPCGF1 after a 9-day treatment with DMSO or 1 μM VTP50469. (H) Bar plot depicting the percentages of CD11b+ cells as shown in panel G. (I). Wright-Giemsa stain of OCI-AML2 cells expressing sgLuc and sgPCGF1, after a 9-day treatment with DMSO or 1 μM VTP50469. Scale bar, 10 μm. (J) Flow cytometry analysis of the percentages of CD11b+ cells in OCI-AML2 cells expressing sgLuc and sgBCOR, after a 9-day treatment with DMSO or 1 μM VTP50469. (K) Bar plot depicting the percentages of CD11b+ cells as shown in panel J. (L) Wright-Giemsa stain of OCI-AML2 cells expressing sgLuc and sgBCOR, after a 9-day treatment with DMSO or 1 μM VTP50469. Scale bar, 10 μm. (M-N) Kaplan-Meier survival curve of mice after transplantation of MV4-11 AML cells expressing the indicated sgRNAs. P values were calculated using the log-rank test. IC50, 50% inhibitory concentration; n.s., not significant; SSC, side scatter; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P <. 0001.
