Figure 2.

NOX1 and p47^phox are required for K-Ras-induced ROS generation and subsequent malignant transformation. (a) Levels of ROS as assessed by DCFDA fluorescence in Rat2 cells or K-Ras^V12-transduced cells after transfection with siRNA targeting NOX1, NOX2, NOX3, NOX4, or scrambled siRNA. (b) Soft agar colony formation of K-Ras-transduced cells after transfection with siRNA targeting NOX1, NOX2, NOX3, NOX4, or scrambled siRNA and control vector MFG-transduced cells. (c) Levels of ROS as assessed by DCFDA fluorescence in K-Ras^V12-transduced cells after transfection with siRNA targeting p47^phox, Noxo1, or scrambled siRNA. (d) Soft agar colony formation of K-Ras^V12-transduced cells after transfection with siRNA targeting p47^phox, Noxo1, or scrambled siRNA and control vector MFG-transduced cells. (e) Western blot analysis for p47^phox after immunoprecipitation with anti-phospho-serine, -phospho-threonine or -phospho-tyrosine antibody or particulate after separation of cytosol and particulate. Error bars represent mean±S.D. of triplicate samples. *P<0.001