Figure 1.

(A) Cell death was measured 48 to 144 h upon exposure to 50 µM TMZ by flow cytometry using PI staining in LN-229 cells. (B) Cell death was measured 120 and 144 h upon exposure to 50 µM TMZ by flow cytometry using Annexin V/PI staining in LN-229 cells. (C) Proliferation arrest was measured upon exposure up to 100 µM TMZ by colony formation assay (CFA). (D) Senescence was measured by flow cytometry using C₁₂FDG staining 120 and 144 h after exposure to 50 or 100 µM TMZ. (E) Cell cycle distribution was measured 48 to 144 h upon exposure to 50 µM TMZ by flow cytometry using PI staining in LN-229 cells. (F) Representative histograms showing the appearance of cells harboring > 2n DNA 120 h after TMZ (50 µM) exposure. (A–E) Experiments were performed in triplicate, and differences between treatment and control were statistically analyzed using Student’s t-test (not labeled = not significant, * p < 0.1, ** p < 0.01, *** p < 0.001). Concerning cell cycle distribution, at all the time points, a significant (***) decrease of cells in G1 phase was observed.