Figure 3. Effect of metformin on the level of constitutive expression of γH2AX in TK6 cells. Exponentially growing TK6 cells were untreated (Ctrl) or were grown in the presence of 0.1, 1.0, 5.0 and 10 mM metformin (MF) for 48 h. The expression of γH2AX was detected with phospho-specific (Ser139-P) Ab and cell fluorescence was measured by flow cytometry. Based on differences in DNA content the cells were gated in G1, S and G2M phases of the cell cycle and the mean values of γH2AX IF for cells in each of these cell cycle phases were calculated. The numerical figures show the percent reduction in mean values of γH2AX IF of the metformin-treated cells with respect to the mean values of the untreated cells (Ctrl) in the respective phases of the cell cycle.