Caki 2

In this study, we used two human CCRCC cell lines. Caki-1 and Caki-2 cells were purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea). The Caki-1 and Caki-2 cells were cultured in DMEM medium with 10% fetal bovine serum (Gibco BRL, Gaithersburg, MD) and streptomycin and penicillin (100 U/ml) in 5% CO₂ and 37°C.

Renal cancer cell lines (Caki-1, Caki-2, A498) were obtained from the Korean Cell Line Bank (Seoul, Korea) and American Type Culture Collection (Manassas, VA, USA), and cultured in McCoy’s 5A and RPMI1640 supplemented with 10% fetal bovine serum. VHL-null RCC4 cell lines were kind gift from Dr. Jong-Wan Park [38 (link)], and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum. Cells were incubated in a humidified atmosphere at 37 °C under normoxia (20% O₂ and 5% CO₂) or hypoxia (1% O₂ and 5% CO₂). Cell viability assay was performed using crystal violet staining [39 (link)]. Briefly, cells were cultured in 24-well tissue culture dishes with or without drug treatment. After incubation with drugs, washed cells were fixed using 4% paraformaldehyde, and then stained with 0.5% crystal violet solution for 20 min at room temperature. Optical density of crystal violet was analyzed using 1% SDS solution and measured by an absorbance reader (BioTek, Winooski, VT, USA) (OD570).

The human RCC cell lines used in this study, ACHN, A498, A704, Caki-1, Caki-2, SN12C, SNU-1272, and SNU4600, were purchased from the Korean Cell Line Bank (Seoul, Korea). ACHN, A498, A704, Caki-1, Caki-2, and SN12C cell lines were cultured in Dulbecco’s modified Eagle medium supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 10 µg/ml gentamicin (Gibco). SNU-1272 and SNU-4600 cell lines were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated FBS and 10 µg/ml gentamicin.