Fig. 5.

Inhibition of Akt and FoxO3a phosphorylation plays a crucial role in the elevation of Bim expression. (A) Bim transcriptional level was assessed by RT‐PCR in PC‐9GROR and H1975OR cell with the indicated treatment. (B) The Akt and Erk phosphorylation proteins were measured by western blot assay combined treated with aspirin and osimertinib or not in PC‐9GROR and H1975‐OR cells. GAPDH was used to confirm equal gel loading. (C) The FoxO3a phosphorylation was measured by western blot assay combined treated with aspirin and osimertinib or not in PC‐9GROR and H1975‐OR cells. GAPDH was used to confirm equal gel loading. GAPDH was used to confirm equal gel loading. (D) Immunofluorescence staining showed the subcellular localization of FoxO3a in PC‐9GROR and H1975‐OR cells with indicated treatments. (E) Cell viability of PC‐9GROR and H1975‐OR cells treated with osimertinib + aspirin + IGF‐1 (200 ng·mL⁻¹) for 48 h was assessed by the MTT assay. All the experiments were measured in triplicate.