Research Paper Volume 13, Issue 10 pp 14289—14303

Autophagy deficiency downregulates O6methylguanine-DNA methyltransferase and increases chemosensitivity of liver cancer cells

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).

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).