Cell cycle arrest in G1 and G2 in response to EGF removal. (A): Capan-2 spheroids control or 6 days after EGF removal. Visualization of EdU incorporation (left), Fucci-red (middle) and Fucci-green (right) expressing cells. Nuclei are stained with DAPI (blue). EdU has been incorporated for 24 h. Scale bar: 50 μm. (B) Quantification of the percentage of EdU (top), Fucci-red (middle) and Fucci-green (bottom) expressing cells as a function of the distance to the spheroid surface. 0-40 μm, 40-80 μm and 80-120 μm intervals are considered. For each condition, data correspond to the mean+/−SEM of percentage of positive cells on 10-30 sections from 2 or 3 independent experiments with 5 to 10 different spheroids each.