Figure 5.

Myosin IIA knockout by clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRISPR/Cas9) attenuates H₂O₂-induced morphological changes and apoptosis in PC12 cells. (A) Stable Myh9 knockout PC12 cells were established by CRISPR/Cas9. Cell lysates from single cell colonies #1, #2, #3 and wild-type (WT) cells were analyzed by western blotting using myosin IIA, myosin IIB and GAPDH antibodies. (B) The WT and Myh9 knockout PC12 cells were untreated or treated with 100 μM H₂O₂ for 12 h. Cells were imaged using phase-contrast microscopy. Bar, 10 μm. (C) Viability of WT and Myh9 knockout PC12 cells treated or untreated with H₂O₂ was evaluated by MTT assay. (D) Caspase-3 activity was measured by caspase-3 activity assay kit. Values were represented as mean ± SD from three independent experiments (^#P < 0.05, ^##P < 0.01).