Figure 3.

Localization of myosin IIB 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 IIB (green), F-actin (red) and DAPI (blue). Neurons untreated (C) or treated (D) with 100 μM H₂O₂ for 12 h were stained with myosin IIB (green), F-actin (red) and DAPI (blue). Images were obtained by confocal microscopy. Bar, 5 μm. The co-localization of myosin IIB 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). (G) Protein interaction between myosin IIB and F-actin was determined by co-immunoprecipitation. Following treatment, cell lysates were immunoprecipitated with anti-actin antibody and isotype-matched IgG control. Each precipitated sample was detected for the presence of myosin IIB and actin by immunoblot analysis using specific antibodies. WCL prior to the immunoprecipitation served as input controls. (H) Expression of Myosin IIB in neurons treated or untreated with H₂O₂ was evaluated by immunoblot analysis.