Figure 7.

Isotope-coded affinity tags identify p21-activated kinase 2 (Pak2) as a mediator of oxidative signaling. A: Biotin-iodoacetamide (BIAM) labeling analysis of Pak2 from SK-CO-15 cells treated with 1 mmol/L H₂O₂ for 15 minutes. Immunoblot analysis was used to detect the levels of total Pak2. Immunoblot shown is representative of at least three replicate experiments. B: Immunofluorescence analysis of SK-CO-15 cells treated with the Pak2 inhibitor FRAX597 at 2 μmol/L for 1 hour, followed by exposure to 1 mmol/L H₂O₂ for 15 minutes. Analysis detected phosphorylated Crk-associated substrate (p-Cas; red), and total focal adhesion kinase (FAK; green) localization in nuclei was stained with DAPI (blue). Scale bars = 20 μm (B). CTL, control.