Human RCC cell lines (Caki-2, 786-O, A498, A704, 769-P, ACHN, RCC4+vec, and RCC4+VHL) were maintained in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% FBS, penicillin (24 U/ml), and streptomycin (25 μg/ml). Caki-2 (2.0 × 104 cells per well), 786-O (1.0 × 104 cells per well), A498 (1.0 × 104 cells per well), A704 (2.0 × 104 cells per well), 769-P (1.0 × 104 cells per well), ACHN (1.0 × 104 cells per well), RCC4+vec (0.7 × 104 cells per well), and RCC4+VHL (0.8 × 104 cells per well) cells were plated into 96-well plates 1 day before GO289 treatments. Two days after treatment, cells were stained with Hoechst 33342 (2 μg/ml; Invitrogen) for 1 hour and counted with ArrayScan XTI (Thermo Scientific).

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