Rpmi 1640 culture media supplement

The human renal cell carcinoma cell lines A498, Caki-1, ACHN and 786-o were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Normal human primary kidney tubular epithelial (HK-2) cells were obtained from Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All cancer cells were cultured in RPMI 1640 culture media supplement (Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (FBS, Sigma, St. Louis, MO, USA). NHK cells were grown in Dulbecco's Modified Eagle's medium (Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum. All the cells were cultured at 37 °C in a humidified atmosphere of 5% CO₂. OSU-T315 (CAS 1333146-24-9, ILK inhibitor, CA, USA) were prepared in dimethyl sulfoxide (DMSO) and stored at ‑20˚C. A498 and ACHN cells were seeded at 1x10⁶ cells per 60‑mm dish and incubated at 37˚C for 16 h. Inhibitors (0.1% of culture medium) were then added to culture media (OSU-T315, 0.1 µM), and cells were incubated at 37˚C for 48 h.