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. Author manuscript; available in PMC: 2020 Apr 8.

Published in final edited form as: Radiat Res. 2018 Aug 22;190(5):538–557. doi: 10.1667/RR15099.1

FIG. 3. — Influence of chloroquine (CQ) on radiation sensitivity in H460wt and H460crp53 cells. Panel A: Inhibition of autophagy by CQ. Fluorescence microscopy showing acridine orange-stained vacuoles induced by 6 Gy radiation alone or with CQ (10 μM) treatment (scale bar = 200 μm). Panel B: Inhibition of autophagy by CQ. Western blot showing autophagy blockade by CQ (10 μM) based on levels of p62/SQSTMQ and LC3 II. Cells were pretreated with CQ for 3 h prior to irradiation and protein was isolated after 3 days. The bar graph in each panel indicates the relative band intensity generated from densitometric scans of three independent experiments in arbitrary densitometric units. Panel C: Influence of autophagy inhibition on radiation sensitivity. Clonogenic survival assay indicating that CQ (10 μM) increased radiation-induced cell growth inhibition in H460wt cells, but not in H460crp53 cells. Panel D: Influence of autophagy inhibition on radiosensitivity. Cell viability assay indicating that CQ increased sensitivity of H460wt cells to radiation (6 Gy), but not H460crp53 cells. Panel E: Influence of autophagy inhibition on radiation-induced apoptosis. Annexin V/PI staining indicating that CQ (10 μM) increased radiation-induced apoptosis (after 2 days) in H460wt cells, but not in H460crp53 cells. Results were from three independent experiments. *P < 0.05, irradiated only cells vs. irradiated and CQ-treated cells.

FIG. 3. — Influence of chloroquine (CQ) on radiation sensitivity in H460wt and H460crp53 cells. Panel A: Inhibition of autophagy by CQ. Fluorescence microscopy showing acridine orange-stained vacuoles induced by 6 Gy radiation alone or with CQ (10 μM) treatment (scale bar = 200 μm). Panel B: Inhibition of autophagy by CQ. Western blot showing autophagy blockade by CQ (10 μM) based on levels of p62/SQSTMQ and LC3 II. Cells were pretreated with CQ for 3 h prior to irradiation and protein was isolated after 3 days. The bar graph in each panel indicates the relative band intensity generated from densitometric scans of three independent experiments in arbitrary densitometric units. Panel C: Influence of autophagy inhibition on radiation sensitivity. Clonogenic survival assay indicating that CQ (10 μM) increased radiation-induced cell growth inhibition in H460wt cells, but not in H460crp53 cells. Panel D: Influence of autophagy inhibition on radiosensitivity. Cell viability assay indicating that CQ increased sensitivity of H460wt cells to radiation (6 Gy), but not H460crp53 cells. Panel E: Influence of autophagy inhibition on radiation-induced apoptosis. Annexin V/PI staining indicating that CQ (10 μM) increased radiation-induced apoptosis (after 2 days) in H460wt cells, but not in H460crp53 cells. Results were from three independent experiments. *P < 0.05, irradiated only cells vs. irradiated and CQ-treated cells.