Fig. 2

Pulsatile, high concentration of sunitinib leads to increase in the sub-G1 population, activation of caspase-3/7 and upregulation of autophagic flux. a FACS analysis utilizing PI staining of 786-O RCC cells exposed to 20 μM of sunitinib after 1, 3, 9 h or 15 h after the 9-h exposure (9 > 24). Sunitinib leads to proportional increase in sub-G1 population, indicative of cell death. b 786-O RCC cells were exposed for 3, 6, 9 and 24 h to 5 or 20 μM of sunitinib and activation of caspase-3/7 was measured. Data presented relative to control = 1. c, e LC3BI, LC3BII and p62 protein levels in control and treated with the indicated concentrations of sunitinib cell lysates were analyzed by Western blotting. d, f Quantification of LC3BII/LC3BI conversion ratio and p62 expression, respectively. For both figures, protein level was initially normalized towards the loading control, β-actin, and then towards the corresponding control- untreated sample