Figure 7. Cells with loss-of-function alleles for TP53 or BAX have altered sensitivities to small molecule inhibitors of various signaling pathways.

A. MOLM-13 and MV4;11 cells, transduced with indicated sgRNAs/Cas9 viruses, were screened with a panel of inhibitors targeting various molecular pathways and assessed using the drug screening method described in Figure 2A. Fold changes in IC₅₀ values, (log10 scale) for those inhibitors concordant across both cells lines relative to nontargeting controls are shown. B. Analyses of FLT3 inhibitor sensitivities, in MOLM-13 and MV4;11 parental and KO cells, as indicated. Drug sensitivity was measured as described in Figure 2A, with IC₅₀ estimated from triplicates. C. Histogram of sensitivities (IC₅₀ values) to a series of NTRK inhibitors in MOLM-13 cells, transduced with indicated sgRNAs/Cas9 viruses. D. Drug sensitivity to NTRK inhibitors entrectinib (top) and larotrectinib (bottom) as measured in Figure 2A and 2F, in MOLM-13 cells transduced with indicated sgRNAs/Cas9 viruses. E. Expression levels of NTRK1, 2, and 3 mRNA using qRT-PCR analysis with RNAs isolated from MOLM-13 and MV4;11 cells with TP53 KO alleles or non-targeting (NT) control. Expression values were normalized to 36B4 gene expression levels. Statistical significance was determined ANOVA with Tukey posttest (**, p<0.01; ***, p<0.001). F. Western blot analyses of proteins extracted from MOLM-13 parental and MOLM-13 transduced with TP53 sgRNA/Cas9 viruses and treated overnight with venetoclax (Ven, 100 nM), entrectinib (Entr, 100 nM) or vehicle (DMSO), and identified with antisera to pan NTRK, phosphorylated NTRK (Tyr516), phosphorylated ERK1/2 (Thr202/Tyr204), ERK1/2, phosphorylated AKT(Thr308), AKT and GAPDH. G. Western blot analyses of proteins from AML patient samples with known TP53 mutant (TP53^mut) or WT (TP53^wt) status. Samples 17-00248 and 15-00309 have FLT3-ITD mutations; sample 17-00300 has a FLT3^D835E mutation.