Figure 1. Loss of autophagy increases DNA damage. (A) Western analysis of Atg5 in cells infected with either Atg5-shRNA1/shRNA2 lentivirus or empty lentivirus control vector; GAPDH was used as a loading control. (B) Western analysis of γ-H2AX protein levels in epirubicin-treated (2 μM; 24 h) cells. (C) Accumulation of γ-H2AX analyzed by immunofluorescence as foci formation in epirubicin (4 μM; 6 h) treated cells (scale bar, 100μm). (D) Quantification of γ-H2AX+ cells. Data are expressed as mean±SD. ** P≤0.01. (E) Wild-type, vector control, and sh-Atg5 cells were treated with 4 μM epirubicin for 12 h, then cells were analyzed by comet assay (scale bar, 20μm).