Extended Data Fig. 6: Lentiviral expression of MEN1 mutants confers resistance in cell lines.
From: MEN1 mutations mediate clinical resistance to menin inhibition
a) Western blot in MOLM13 cells showing expression of HA-tagged MEN1-WT and M327I-mutant construct. Representative Western Blot of 3 independent replicates. b) Dose-response curves of MOLM13 cells to revumenib upon expression of MEN1 mutants compared to -WT. Cell counts were measured by flow cytometry and displayed relative to the DMSO control (mean +/− SEM, n = 4, each 3 technical replicates). c) Dose-response curves of MV4;11 and OCI-AML3 cells to revumenib upon expression of MEN1 mutants or -WT measured by Cell-titerGlo (mean +/− SD, n = 3). d) Induction of differentiation marker expression by revumenib in OCI-AML3 cells expressing MEN1 mutants compared to WT measured by flow cytometry (MFI CD11b) (n = 3, mean +/− SD). An unpaired, two-tailed t-test was used for statistical analysis. e) Quantification of the cytological assessment for blast morphology by a hematopathologist (n = 5, median, box: 25th to 75th percentile, whiskers: range). One-way ANOVA with correction for multiple comparisons was used for statistical analysis. f) Dose-response curves of OCI-AML3 cells to revumenib (top panel) or MI-3454 (bottom panel) upon expression of MEN1 mutants compared to -WT. Cell counts were measured by flow cytometry and displayed relative to the DMSO (mean +/− SEM, n = 4, each 3 technical replicates). g) Western blot in MV4;11 cells showing expression of HA-tagged MEN1-WT and -mutant constructs. Representative Western Blot of 2 independent replicates. h) Dose-response curves of MV4;11 cells to revumenib upon expression of MEN1 mutants compared to -WT. Cell counts were measured by flow cytometry and displayed relative to the DMSO (mean +/− SEM, n = 4, each 3 technical replicates). b, f, h) Statistical analysis was performed using an unpired t-test, two tailed, multiple comparisons. *** p < 0.001; **p < 0.01; *p < 0.05.
