Differential regulation of key markers of VHL function by type 1 and type 2 pVHL
mutants in 786-O. (A) 786-O cells were stably transfected with empty vector control or wild-type (WT) or mutant pVHL19 expression constructs, as indicated. Cells were grown to confluence on collagen I and prepared cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for VHL, HIF-2α, GLUT-1, α5 integrin, cyclin D1, and α-tubulin. A faster migrating band is seen for del 114–178 upon darker exposure of the VHL blot (not shown). Quantification of the band intensities for the α5 integrin blot (as a percent of the band with greatest intensity) is provided at the bottom. (B) Indicated 786-O stable cell pools were grown to confluence on collagen I and photographed by digital phase microscopy (original magnification of 100×). Note the disorganized morphology of the control and the VHL del 114–178 and RC161/2QW-expressing cells.