Fig. 4.

Deregulation of HIF2α does not fully explain the increased requirement of 786-O VHL−/− cells for CDK6, MAP2K1, and MET. (A) Immunoblot analysis of 786-O cells infected to produced HA-pVHL alone (VHL) or HA-pVHL and a stabilized version of HIF2α (VHL + dP →A). Parental cells infected with an empty virus (vector) served as controls. (B) Percentage loss of viability (% LOV) of cells (normalized to scrambled shRNA) infected as in A after subsequent infection with 5 μl (gray bars) or 10 μl (black bars) of lentiviruses encoding shRNAs targeting CDK6, MET, or MAP2K1. Cells were grown in 96-well plates and were assayed for survival in triplicate using alamarBlue 5 days post-infection. Error bars = 1 SEM.