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. 2022 Jul 1;132(13):e145099. doi: 10.1172/JCI145099

Figure 5. RBM10 deficiency decreases the Bcl-xS to Bcl-xL ratio to limit the apoptotic response to EGFR TKI therapy.

Figure 5

(A) RBM10 regulates Bcl-x mRNA splicing into Bcl-xS (proapoptotic) and Bcl-xL (antiapoptotic) isoforms. (B and C) qRT-PCR analysis of the Bcl-xS to Bcl-xL ratio (mRNA levels) in H3255 (B) and PC-9 (C) cells expressing shRBM10 or shScr control. Data are shown as the mean ± SEM of the FC after normalization to the housekeeping gene (GAPDH). (D and E) Conventional PCR analysis using validated primers to detect both Bcl-xL and Bcl-xS isoforms in H3255 and PC-9 cells expressing either shScr control or shRBM10 with or without genetic rescue of Bcl-xS. (F and G) H3255 and PC-9 (EGFR L858R and EGFR del19, respectively; WT RBM10) cells treated with osimertinib for 48 and 72 hours, which express either shRBM10 or shScr control paired with or without genetic rescue of Bcl-xS. Cell lysates were harvested, and expression of the indicated proteins was measured by Western blotting. (H and I) Caspase 3/-7 activity was measured using the Caspase-Glo 3/7 assay. Each bar represents the mean ± SEM of the FC after normalization to the DMSO control. Data represent 3 independent experiments. *P < 0.05, by 1-way ANOVA.