Additional RCC10 cell lines were generated. (A) RCC10 VHL-null cells were stably transfected with pVHL30 expression constructs. Cells were grown to confluence and prepared cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for VHL, HIF-2α, and GLUT-1. α-tubulin was also assayed to demonstrate equal loading. (B) RCC10 VHL-null cells were stably infected with retroviruses containing mutant pVHL30 expression constructs (S65W, N78S, L158P) and assayed along with original control and WT VHL RCC10 cells as in (B).