Figure 2.

LCC does not induce apoptosis in C6 cell but depletes cells at G2 and M phases. A-D) The effect of CHX on cell cycle profile was examined by flow cytometry analysis. C6 cells were seeded in 10 cm dishes at a density of 1.0 × 10⁶ per dish. After treatment with CHX at indicated concentration for 24 h, cells were trypsinized, washed twice by PBS, and fixed with ice-cold 70% ethanol. DNA content was measured after stained by PI solution [0.1% sodium citrate (w/v), 0.1% Triton X-100 (v/v), 50 mg/ml PI in demonized/distilled water]. Percentages of G2/M (G2) phase cells were indicated. E) Western immunoblotting analysis on molecules that are regulators and indicators of cell stress and cell death. Experiments were done as in Figure 1D.