GFP-WIPI-1 puncta formation upon serum and amino acid starvation. GFP-WIPI-1 U2OS cells were treated with nutrient-rich culture medium (DMEM/FCS), serum-free culture medium (DMEM), or with medium lacking both serum and amino acids (EBSS) for 0.5, 1, and 2 h. Fluorescence images were automatically acquired and 2 h treatment images are shown ((a)–(c)). The number of GFP-WIPI-1 puncta-positive cells (d), and of GFP-WIPI-1 puncta per cell (e) was automatically determined. Each measure point represents mean value from up to 3000 individually analyzed cells per treatment condition ± SD (, each in triplicates). Scale bars: 20 μm.
