Extended Data Fig. 8: MS177 exhibits potent effect on inducing leukaemia cell growth inhibition, apoptosis, and cell cycle progression arrest.
(a) Immunoblotting for EZH2 and GAPDH in four MLL-r leukaemia cell lines, treated with the indicated compound (0.5 µM) for 24 hours. (b) Effect of a 24-hour treatment with different concentrations of MS177 on proliferation of the two indicated primary samples from de-identified AML patients. Y-axis shows mean ± SD after normalization to DMSO-treated (n = 3). (c) Proliferation of primary AML cells treated with 1 µM of C24 or MS177, relative to DMSO, for 48 hours (n = 3; mean ± SD; unpaired two-tailed Student’s t-test). (d-e) Immunoblotting for EZH2, PRC2 subunits and cMyc (d) and RT-qPCR for EZH2:H3K27me3-cobound genes (e) in primary AML cells, treated with the indicated compound (0.5 µM) for 24 hours. For e, y-axis shows RT-qPCR signals after normalization to those of GAPDH and to DMSO-treated cells (n = 3; mean ± SD; unpaired two-tailed Student’s t-test). (f) Growth of K562 cells treated with MS177, relative to DMSO, for the indicated time. Y-axis shows mean ± SD after normalization to DMSO-treated (n = 3). (g) Representative view of colonies formed by murine HSPCs in the presence of DMSO or MS177. (h-i) Immunoblotting for EZH2 and IKZF1/3 (h) and growth (i) of EOL-1 cells after the indicated treatment of compound (0.5 µM for all). Pom, pomalidomide. For i, y-axis shows relative growth after normalization to DMSO-treated (n = 3; mean ± SD; unpaired two-tailed Student’s t-test). (j) Representative flow cytometry-based histograms showing the DNA content in MOLM-13 cells, treated with indicated compound for 24 hours. (k-l) Immunoblotting for apoptotic markers in EOL-1 (k) or MV4;11 (l) cells after the indicated compound treatment. (m) Proliferation of the EZH2-depleted (sgEZH2) or control (sgEV) MOLM-13 cells, treated with DMSO or MS177 (0.1, 0.5 or 1 µM) for 48 hours (n = 3; mean ± SD; unpaired two-tailed Student’s t-test). *, **, and *** denote the P value of < 0.05, 0.01 and 0.005, respectively. NS denotes not significant. Numerical source data, statistics, exact P values and unprocessed blots are available as source data.
