FIG. 2.

Mitochondrial oxidative stress activates PKD at the mitochondria. (A to C) Cells were stimulated with H₂O₂ (10 μM, 10 min), DPI (20 μM), or rotenone (10 μM) as indicated. Mitochondrial fractions were prepared, and lysates were resolved by SDS-PAGE and immunoblotted against the indicated proteins. All results are typical of three independent experiments. (D) Cells were stimulated with H₂O₂ (10 μM), DPI (20 μM), or rotenone (10 μM) for the indicated times. PKD was immunoprecipitated (IP), and a PKD substrate kinase assay was performed. Expression of PKD was determined by immunoblotting with anti-PKD (α-PKD; bottom panels).