Figure 7.

Caspase, ROCK or myosin II inhibition attenuates H₂O₂-induced neuronal apoptosis, membrane blebbing and neurite retraction. PC12 cells or neurons were untreated or pretreated with 1 μM blebbistatin, 10 μM Y27632 or 10 μM z-VAD-fmk for 1 h prior to 100 μM H₂O₂ treatment for 12 h. N-acetyl-L-cysteine (NAC; 500 μM) served as positive control. (A) Viability of PC12 cells and neurons was evaluated by MTT assay. (B) Caspase-3 activity of PC12 cells and neurons was determined by caspase-3 activity assay kit. (C) Representative phase-contrast micrographs of PC12 cells. Bar, 10 μm. (D) TEM of treated PC12 cells. Bar, 1 μm. (E) PC12 cells with membrane blebs were counted and the percentage of these cells with respect to the total cell population was calculated. (F) Neurite length was measured using the ImageJ software. The results were represented by the percentage of neurite length compared to control. Results were expressed as mean ± SD from three independent experiments (^##P < 0.01 vs. control, *P < 0.05 vs. H₂O₂-treated cells, **P < 0.01 vs. H₂O₂-treated cells).