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Figure 4


Fig. 4. Effect of mitochondria-generated ROS on RhoA activity and protein levels in A549 cells during hypoxia. (A) A549 cells were infected with null adenovirus (Sham) or adenovirus expressing Myc-tagged GPX1 (AdGPx) and subsequently exposed to 21% O2 or 1.5% O2 for 15 minutes. 1% Triton X-100-soluble membrane fractions were obtained and RhoA translocation was evaluated by western blot. Mean ± s.e.m. (n=3), **P<0.01. A representative western blot is shown for RhoA and Myc. (B) WT and ({rho}0)-A549 cells were exposed to 21% O2 or 1.5% O2 for 15 minutes and then processed as described above. Mean ± s.e.m. (n=3), **P<0.01. (C) WT and ({rho}0)-A549 cells were exposed to 21% O2 or 1.5% O2 for 15 minutes and cell lysates were subjected to a pull-down assay with a GST-tagged fusion protein RBD and processed as described. Mean ± s.e.m. (n=3), **P<0.01. (D) A549 cells stably transfected with RNAi against Drosophila HIF (dHIF) or the Rieske iron sulphur protein (FeS) were exposed to 21% O2 or 1.5% O2 for 15 minutes and then processed as described above. Mean ± s.e.m. (n=3), **P<0.01.





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