Genetic manipulations of hTERT expression alter the responses of EGFRm NSCLC cell lines to osimertinib. (A–F) The indicated cell lines transfected with scrambled control or hTERT siRNA for 48 h (A–C) or expressing pLKO.1 or shTERT (D and E) were exposed to DMSO or 200 nM osimertinib (Osim) for 24 h (A and D), 48 h (B and E), or 72 h (C and F). (G–I) The indicated cell lines expressing vector (V) or hTERT gene were exposed to DMSO or 100 nM osimertinib for 24 h (G), 48 h (H), or 72 h (I). The proteins of interest were detected with western blotting (A, D, and G). Annexin V-positive cells were determined with flow cytometry (B, E, and H). Cell numbers were estimated with the SRB assay (C, F, and I). The data are means ± SDs of triplicate (B, E, and H) or four replicate (C, F, and I) determinations. Statistical analysis was conducted with one-way ANOVA test (B) or two-sided unpaired Student’s t test (E and H). CF, cleaved form. Source data are available for this figure: SourceData F4.