Fig. 5

Effect of kidney cancer spheroid number on AKT signaling. p-AKT (Ser 473) analysis in 786-O (A-B) or WT-CLS1 (C-D) spheroids stimulated or not with collagen I. Representative p-AKT immunoblots obtained with total protein extracts from 786-O (upper A) or WT-CLS1 (upper C) spheroids. p-AKT is shown as a 60 kDa protein. Lower A and C, representative Ponceau S staining of the membrane. For each independent experiment, band intensities were quantified using Fiji software (B and D) and normalized to the corresponding Ponceau S staining bands seen on the membrane. Values for a single spheroid were arbitrarily set to 1. Data are presented as the mean +/- SEM of 5 to 6 independent experiments, (n = 6 for 786-O and n = 5 for WT-CLS1 spheroid cultures), *p < 0.05, Kruskal-Wallis test, followed by Dunn’s Multiple Comparison Test