FIGURE 7.

Genotoxic effect of EcA and variants in MV4:11 cells. A, comet assay, and B, micronuclei formation of wild-type EcA- and W66Y-treated MV4:11 cells. MV4:11 cells were cultured in complete medium for 24 h, then treated with 0.8 units/ml EcA and variants for 96 h, and analyzed for DNA damage by alkaline Comet assay. The cells were stained with 0.5 μg/ml propidium iodide and observed using a fluorescence microscope (Nikon, Japan). Micronuclei formation was checked by PI staining. Cells were arrested at the cytokinesis stage by cytochalasin-B (0.5 mg/ml) and stained with propidium iodide stain. 1,000 cells were analyzed to check the frequency of micronuclei formation. Images were captured by Olympus BX61 fluorescent microscope and analyzed by Cytovision7.2 software. Untreated (control) cells were taken as negative control. Each experiment was performed three times.