Fig. 2

Nutlin-3a disrupts the fusion of autophagosomes and lysosomes in KRAS MT/p53 WT NSCLC cells. A Downregulation of the mTOR pathway detected by western blotting after nutlin-3a treatment. B Western blotting for the expression of autophagy markers performed after co-treatment of cells with bafilomycin A1 (100 nM) for 1 h in the presence or absence of nutlin-3a for 24 h. C Autophagy flux after cell treatment with nutlin-3a (30 μM) for 24 h after transfection of the mRFP-GFP-LC3 plasmid. Live cell imaging was obtained using a confocal laser scanning microscope (left), and number of puncta per cell were quantified from representative images (right) (n ≥ 21). D Western blotting for the expression of surrogate markers of the autophagic process, p-AMPKα, and p-ACC, performed after nutlin-3a (30 μM) for 24 h. Scale bar: 10 μm