Figure 2. GNE-477 induces apoptosis activation in primary human RCC cells. The primary human RCC cells (“RCC1/RCC2/RCC3”), HK-2 renal epithelial cells (“HK-2”) or the primary human renal epithelial cells (“Epi”) were treated with GNE-477 (50 nM) or the vehicle control (“Veh”, 0.1% DMSO), cells were further cultured for designated time periods (24-48h), and cell apoptosis tested by the mentioned assays (A–E, H, I). Alternatively, RCC1 cells were pretreated for 1h with applied caspase inhibitors (each at 50 μM), followed by GNE-477 (50 nM) stimulation, cells were further cultured for 48-72h, with cell apoptosis and viability examined by nuclear TUNEL staining (F) and CCK-8 (G) assays, respectively. Bars stand for mean ± standard deviation (S.D.). For each assay, n=5. ** p < 0.01 vs. “Veh” cells (A, B, D, E, H, I). ##p < 0.01 vs. “DMSO”-pretreated cells (F, G). Experiments in this figure were repeated five times, and similar results obtained. Scale bar= 200 μm (E).