Figure 1. HR and MMR protein and mRNA expression is reduced in VHL-deficient renal carcinoma cells.

(A, B) Western blotting was performed to measure FANCD2 protein expression in 786-O^VHL–/– and 786-O+VHL^WT cells (A) or RCC4^VHL–/– and RCC4+VHL^WT cells (B) exposed to normoxia or hypoxia (<0.01% O₂) for 24 or 48 h. Vinculin is presented as loading control and numbers below the blots indicate FANCD2 band density relative to vinculin and normalized to 786-O^VHL–/– or RCC4^VHL–/– normoxia samples. (C) Western blotting for additional DNA repair proteins was performed in untreated 786-O and RCC4 matched pair cells. Vinculin is presented as loading control and numbers below the blots indicate band density relative to vinculin and normalized to VHL^WT samples. (D) qRT-PCR was performed to measure FANCD2, BRCA1, RAD51, and MLH1 mRNA levels in untreated 786-O and RCC4 matched pair cells. Expression levels were normalized to 18S rRNA expression and presented as relative to VHL^WT samples. Columns, mean of 3 replicates; bars, SEM.