Figure 6.

Fip200–/– MEFs display increased sensitivity to apoptosis triggered by 3-methyladenine or SBI-0206965, but not to MHY1485, Cpd18, SAR405 or Spautin-1. (A) Protein extracts from Fip200–/– and Fip200+/+ MEFs cultured in full media or Hank’s buffer without glucose (starvation) for 6h, either in the presence (+) or absence (–) of BafA₁, were analyzed by western blot. Autophagy was evaluated by LC3-II and p62. Intensity of these bands, normalized to the loading of each lane and referred to the values of these proteins in Fip200+/+ maintained in growing medium, was shown. Naphthol blue (NB) stained membrane served as a loading control. The image belongs to a representative image out of three independent experiments. (B) Fip200–/– and Fip200+/+ MEFs were challenged with the drugs at the concentrations indicated in the panel. After staining with PI, the percentage of dead cells was determined by flow cytometry. Bar value is the mean ± SEM (n = 3). Student’s t-test *P < 0.01 (C, D) Effector caspase activity (DEVDase activity) was quantified in arbitrary fluorescent units (a.f.u.) after 24 h of treatment with the compounds indicated in the panel. Bar value is the mean ± SEM (n = 3). Student’s t-test *P < 0.01 and ***P < 0.001.