a, EC50 values of MS177 in the indicated AML cell lines (left) and patient samples (right) after a 4-day treatment (n=3).
b, MOLM-13 cell growth after a 48-hour treatment with different concentration of C24, MS177, MS177N1 or MS177N2, relative to DMSO. Y-axis shows mean ± SD after normalization to DMSO-treated (n = 3).
c-e, Effect of a 48-hour treatment with various concentration of MS177, relative to DMSO, on proliferation of four primary AML samples (c) or paired cells from de-identified patients, either in remission or with diagnosed AML (d-e). Y-axis shows mean ± SD after normalization to mock (n = 3) .
f-g, Immunoblotting for the indicated protein in two primary AML samples, treated with DMSO, MS177 or C24 for 24 (f) and 36 (g) hours.
h-j, H3K27me3 immunoblotting (h) and growth (i-j) of EOL-1 cells, treated with the indicated concentration of various EZH2 catalytic inhibitors (h-i), MS177 (i) or UNC6852 (j) for 48 hours (n=3; mean ± SD; unpaired two-tailed Student’s t-test).
k, Colony formation by murine hematopoietic stem/progenitor cells (HSPCs) in the presence of DMSO or MS177 (n=2; mean ± SD).
l-m, Immunoblotting of the indicated protein (l) and growth (m) of MV4;11 cells, treated with 2.5 µM of DMSO, MS177, MS177N2, C24, pomalidomide (Pom), or C24 plus Pom. Y-axis in k shows mean ± SD after normalization to DMSO-treated (n = 3; unpaired two-tailed Student’s t-test).
n-o, Representative image (n) and quantification of colony formation (o; mean ± SD of two experiments) using MV4;11 cells treated with the indicated compound.
p, MOLM-13 cell cycle analysis after a 24-hour treatment with the indicated compound.
q-r, Immunoblotting for various apoptotic markers post-treatment of AML patient cells (q), or the mock-treated (sgEV) or EZH2-depleted (sgEZH2) MOLM-13 cells (r), with the indicated compound for 24 hours.
*, **, 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.