Figure 5. Knockout of MFN2 sensitizes AML cells to BH3-mimetics.

a. Competition-based viability assays in MOLM-13 parental cells treated with MCL1i (left) or MCL1i+Ven (right). Y-axis denotes the GFP⁺ percentage relative to day 0 of treatment (n=3, mean ± SD).

b. Annexin V staining in MOLM-13 cells transduced with sgRNAs targeting MFN2 or control (Rosa), after treatment with BH3 mimetics or DMSO for 24 hrs (n=3, mean SEM, unpaired two-tailed t-test).

c. Competition-based viability assays in MOLM-13 MR cells treated with MCL1i (left) or MCL1i+Ven (right). Y-axis denotes the GFP⁺ percentage relative to day 0 of treatment.

d. Annexin V staining in Kasumi-1 resistant cells transduced with sgRNAs targeting MFN2 or control (Rosa), after treatment with 600 nM AZD5991 or DMSO for 24 hrs (n=3, mean ± SD). Statistics were calculated using two-way ANOVA.

e. Competition-based viability assays in MOLM-13 MR cells transduced with sgRNAs targeting MFN1, MFN2 or control (Rosa) treated with DMSO, AMG176 (MCL1i) or AMG176 (MCL1i)+Ven. Y-axis denotes the GFP⁺ percentage relative to day 0 of treatment (n=3, mean ± SD).

f. Schematic outline of the in vivo experiment validating the synergistic effects of MFN2 ablation and AZD5991 treatment using MOLM-13 xenografts.

g. Bioluminescence images of the MOLM-13 leukemia recipient mice described in (f). The same mice are depicted at each time point (n=5 mice per group). Days post-drug administration are shown.

h. Quantification of bioluminescence emitted from the whole body of each mouse described in (f) at the indicated time points (mean ± SD).

i. Kaplan-Meier survival curves of the MOLM-13 leukemia recipient mice described in (f). X-axis denotes days post-transplantation. The p-values were determined using the Log-rank Mantel-Cox test.

j. Graph depicting changes in the animals’ body weight throughout the experiment described in (f).