Fig. 4

Nutlin-3a induces methuosis-like death of KRAS MT/p53 WT NSCLC cells. Cells were treated with nutlin-3a (30 μM) with or without indicated inhibitors for 24 h. A Cleavage forms of caspase-9, caspase-3, and PARP did not detected by western blotting. Etoposide (ETO) treatment (100 μM) for 24 h was used as a positive control for inducing caspase cleavage. B-I Representative light microscope images are shown. Cell proliferation was determined via cell counting assay. Cells were co-treated with Z-VAD (OMe)-FMK (50 μM) (B), ferrostatin-1 (1 μM) (C), necrostatin-1 (10 μM) (D), or EHT1864 (10 μM) (H) or pretreated with EIPA (5 μM) for 1 h (E–G) or bafilomycin A1 (5 nM) for 2 h (I) before nutlin-3a treatment. Scale bar: 100 μm for b, and 200 μm for e. *p < 0.05, **p < 0.01, ***p < 0.001 compared with control