Figure 7 from N-Myristoytransferase Inhibition Causes Mitochondrial Iron Overload and Parthanatos in TIM17A-Dependent Aggressive Lung Carcinoma

Dependency on TIM17A is a determinant of NMTi sensitivity in lung carcinoma cells. A, TIM17A immunoblotting in the indicated cells treated with 1 μmol/L PCLX-001 (NMTi) for 72 hours. Numbers, band intensity normalized to actin control. B, Boxplot showing CRISPR DepMap Score for LUAD cells (y-axis) estimated using the Chronos algorithm. Negative values suggest decreased cell viability. (KL/K)MUT (HCC44, H460, and H1792) and (KL/K)WT (H522, H1650, and H1437) lung carcinoma cells used in our study are highlighted. C, TIM17A CRISPR Chronos scores (y-axis) plotted against NMTi IC₅₀ (x-axis; Spearman’s Correlation, P = 0.006, ⍴ = −0.43). Axis scales are log-transformed, and the best-fit line (blue) was calculated using a linear model. Gray shading, standard error of the estimate. H460 was added manually based on our calculated IC₅₀ and the publicly available dependency score. D, Cell viability (crystal violet staining) in (KL/K)MUT H1792 and (KL/K)WT H522 lung carcinoma cells transfected with a TIM17-targeting siRNA pool or a non-targeting control. Bar: average; error bars, SEM. One representative experiment from three independent experiments with similar results. ***, P = 0.0009 (Student t test). Bottom, TIM17A immunoblotting of the samples above. E, Colony assays of (KL/K)MUT H460 and (KL/K)WT H522 expressing Tet-inducible TIM17A or control non-targeting shRNA growing in the presence or absence of Dox. Top, Representative images. Bottom, TIM17A immunoblotting of lysates from the cells used for colony assays. GAPDH, loading control.