Figure 2.

Localization of myosin IIA and F-actin in PC12 cells or neurons upon H₂O₂ treatment. PC12 cells untreated (A) or treated (B) with 100 μM H₂O₂ for 12 h were stained with myosin IIA (green), F-actin (red) and DAPI (blue). Neurons untreated (C) or treated (D) with 100 μM H₂O₂ for 12 h were stained with myosin IIA (green), F-actin (red) and DAPI (blue). Images were obtained by confocal microscopy. Bar, 5 μm. The co-localization of myosin IIA with F-actin in PC12 cells (E) or neurons (F) was evaluated on the basis of Manders’ overlap coefficients. Results were expressed as mean ± SD (^##P < 0.01 vs. control). Experiments were performed three times independently. Protein interaction between myosin IIA and actin was determined by co-immunoprecipitation. Following treatment, cell lysates were immunoprecipitated with anti-actin antibody (G) or anti-non-muscle myosin IIA antibody (H). Isotype-matched (IgG) served as negative control (NC). Each precipitated sample was detected for the presence of myosin IIA and actin by immunoblot analysis using specific antibodies. Whole cell lysates (WCL) prior to the immunoprecipitation served as input controls.