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Figure 6


Fig. 6. Dominant negative RhoA prevents hypoxia-induced Na,K-ATPase endocytosis. Permanently transfected A549 cells expressing either vector or dominant negative RhoA (RhoAN19) were grown in the presence of G418. (A) Actin stress fiber formation was assessed in DN-RhoA cells fixed and stained with rhodamin-palloidin and evaluated by using fluorescence microscopy. Representative photomicrographs of control cells (21% O2), and hypoxia (1.5% O2)-treated cells are shown. (B) Na,K-ATPase endocytosis was studied by biotin labeling of surface proteins followed by streptavidin pull-down and western blot analysis using a specific antibody after exposure to 1.5% O2 for 60 minutes. Top: bars represent mean ± s.e.m. (n=3). **P<0.01. Bottom: a representative western blot. (C) Na,K-ATPase endocytosis was studied by biotin labeling of surface proteins followed by streptavidin pull-down and western blot analysis using a specific antibody after treatment with 100 µM H2O2 for 30 minutes. Top: bars represent mean ± s.e.m. (n=3). **P<0.01. Bottom: a representative western blot.





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