Figure 2. Quantification of γH2A.X and total H2A variants by western blot. Whole-cell extracts were prepared from untreated proliferating and bleomycin-treated HCA2 cells (acute DDR (+Bleo, 3 h) and drug-evoked senescence (+Bleo, 5 days)), and subjected to western blot analysis with (A) γH2A.X and (B) generic H2A antibodies. Two exposures (short: 1 min and long: 10 min) are shown for the γH2A.X staining. β-actin was used for normalization purposes. Graphs represent computationally calculated band intensities for each antibody, normalized to β-actin, from western blots repeated for biological triplicates of each condition.