Fig. 6

Gain of function for missense mutant pVHL using adenovirus mediated depletion of UCP. a V155A missense mutant pVHL expressing 786-O stable cell lines were transduced with adenovirus expressing siUCP or siControl at a MOI of 200 and incubated for 48 h. Cells were counted by hematocytometer (n = 3, p < 0.01) (b) At same time cells were harvested and lysed for analysis of UCP-VHL-HIF pathway. Proteins were monitored by immunoblot as indicated. c Cells as indicated were transduced adenovirus expressing UCP-siRNA and/or Cont-siRNA at 200 MOI and then injected into nude mouse subcutaneously (n = 4, p < 0.01). Tumor mass was monitored by 3-4 days during indicated times. d At the end of tumor size measurement, tumors were excised for immunoblot. Pooled tumor pieces of each group were lysed in tissue lysis buffer and immunoblot assay was performed as indicated