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. 2009 Oct 15;284(50):34964–34975. doi: 10.1074/jbc.M109.013771

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Caveolin-1 regulation of hyperoxia-mediated human lung EC ROS production. A, immunoblot analysis of siRNA-treated or untreated HPAECs. Cellular lysates from untransfected (control, no siRNA), scramble siRNA (siRNA that does not target any known human mRNA), caveolin-1 siRNA#1 (Santa Cruz Biotechnology), or caveolin-1 siRNA#2 (ON-TARGETplus siRNA, Dharmacon) transfection were analyzed using immunoblotting with anti-caveolin-1 (a), anti-c-Abl (b), anti-dynamin 2 (c), or anti-actin (d) antibody. B, fluorescent microscopy image of HPAEC ROS production. Lung ECs were treated with either scramble siRNA (Sc) or caveolin-1 siRNA#1 (Santa Cruz Biotechnology), loaded with DCFDA, and exposed to either normoxia or hyperoxia for 3 h. C, graphical representation of data described in Panel B with n = 3 experiments; error bars = ±S.D. *, a statistically significant change (p < 0.05) from control (normoxia, scramble siRNA). **, a statistically significant change (p < 0.05) between scramble siRNA, hyperoxia and caveolin-1 siRNA, hyperoxia. D, fluorescent microscopy image of HPAEC ROS production. Lung ECs were treated with either scramble siRNA (Sc) or caveolin-1 siRNA#2 (ON-TARGETplus siRNA, Dharmacon), loaded with DCFDA and exposed to either normoxia or hyperoxia for 3 h. E, graphical representation of data described in Panel D with n = 3 experiments; error bars = ±S.D. *, a statistically significant change (p < 0.05) from control (normoxia, scramble siRNA). **, a statistically significant change (p < 0.05) between scramble siRNA, hyperoxia and caveolin-1 siRNA, hyperoxia. F, fluorescent microscopy image of HPAEC ROS production. Lung ECs were treated with either scramble siRNA (Sc) or caveolin-1 siRNA#2 (ON-TARGETplus siRNA, Dharmacon), loaded with DHE, and exposed to either normoxia or hyperoxia for 3 h. G, graphical representation of data described in Panel F with n = 3 experiments; error bars = ±S.D. *, a statistically significant change (p < 0.05) from control (normoxia, scramble siRNA). **, a statistically significant change (p < 0.05) between scramble siRNA, hyperoxia and caveolin-1 siRNA, hyperoxia.