Fig. 5

PHD3 depletion leads to impaired ccRCC cell growth and decreases the phosphorylation of p70 S6K and S6 ribosomal proteins. a Western blot analysis of 786-O cells with two siRNA sequences targeting PHD3 under normoxia and hypoxia shows decrease in phosphorylation of p70 S6K and S6 ribosomal protein. Right-hand graphs show phosphorylated (S235/236 and S240/244) pS6 ribosomal protein normalized to total S6 protein. Quantification of two biological replicates, mean ± SEM, fold change to Scr (*p < 0.05, n.s. not significant). b Western blot analysis of RCC4 cells with PHD3 depletion under normoxia and hypoxia decreases the phosphorylation of p70 S6K and S6 ribosomal protein. Quantification of two biological replicates, mean ± SEM, fold change to Scr (*p < 0.05, n.s. not significant). c 786-O and RCC4 3D colony forming in Matrigel®. Quantification of three (786-O) or two (RCC4) biological replicates with triplicate wells for each siRNA treatment, mean ± SEM, fold change to Scr (**p < 0.01, ***p < 0.001). Representative images in the panel acquired by using ×4 objective, scale bar 100 μm