Tight junctions are affected by levels of HIF-2α in 786-O. (A) Parental 786-O cells (786-O), 786-O cells stably transfected with VHL (VHL +), and 786-O cells infected with an empty retrovirus (pSuperRetro) or infected with HIF-2α shRNAs (HIF2α shRNA), as described in , were grown for 1 week past confluence on collagen I coated culture dishes. Cell lysates were prepared, equally loaded, and western blotted for HIF-2α, cyclin D1, p27, α-tubulin and ZO-1 (both upper and lower immunoreactive species that were seen in Figure 9). (B) Indicated cell lines that were assayed in (A) were grown to confluence on coverslips and immunostained for ZO-1 (left panels; original magnification of 1000×). DAPI labeled nuclei of corresponding cells are also shown (right panels). (C) Indicated cell lines that were assayed in (A) and (B) were grown to confluence on collagen I and photographed by digital phase microscopy (original magnification of 100×).