Figure 5.
Resveratrol-promoted increase of LC3-II protein abundance and GFP-LC3 puncta is partially insensitive to wortmannin treatment. Stable GFP-WIPI-1 U2OS cells were treated with control medium (CM), resveratrol (Res), wortmannin (WM), resveratrol plus wortmannin (Res/WM), nutrient-free medium (EBSS) or nutrient-free medium plus wortmannin (EBSS/WM) for 3 h. Bafilomycin A1 (Baf A1) was added after 2 h as indicated (+). LC3-II protein abundance was determined and normalized over tubulin. Each bar represents mean values ± SD (n = 6) (A). Stable GFP-LC3 U2OS cells were treated as above and subjected to automated fluorescence image acquisition (B) and analysis (C). Each bar represents mean values ± SD from the analysis of 1553–3207 individual cells (n = 3). p-values: *p < 0.05; **p < 0.01; ***p < 0.001; ns: not significant. Scale bars: 20 µm.