Renal epithelial basal medium

HK-2 human kidney proximal tubular cells were purchased from American Type Culture Collection and cultured in renal epithelial basal medium (Gibco) with manufacturer provided supplements in an incubator with 5% CO₂ at 37°C.

HK-2 human kidney proximal tubular cells (ATCC, Manassas, VA, USA) were cultured in renal epithelial basal medium (Gibco BRL, Grand Island, NY, USA) with manufacturer-provided supplements. Cells were incubated with 100 mg/mL ioversol or vehicle (PBS) for various times, with PBS or PC at various concentrations (0.2, 1, and 2 ng/mL), and then evaluated morphologically and via Western blotting. To verify the effect of PC on contrast-induced autophagy, five conditions were included: (1) untreated cells, (2) cells treated with ioversol (100 mg/mL), (3) cells treated with PC (1 and 2 ng/mL), (4) cells treated with chloroquine (CQ, 10 and 20 μM), an inhibitor of autophagic flux, and (5) various combinations of these treatments. CQ was applied 4 h prior to the other agents. Cell lysate from each sample was loaded to ensure separation of LC3B-I and -II, and probed with LC3 antibody.