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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. 4. — 3-MA fails to alter radiation sensitivity in H460wt or H460crp53 cells. Panel A: Inhibition of autophagy by 3-MA. Western blot showing autophagy blockade by 3-MA (1 mM) based on levels of LC3 II. Cells were pretreated with 3-MA for 3 h prior to irradiation and protein was collected 3 days postirradiation. The bar graph in each panel indicates the relative band intensity generated from densitometric scans of two independent experiments in arbitrary densitometric units. Panel B: Inhibition of autophagy by 3-MA. Cells were pretreated with 3-MA for 3 h prior to irradiation and cells were stained with acridine orange 3 days postirradiation (scale bar = 200 μm). Panel C: Influence of 3-MA on radiation sensitivity. Cell viability assay indicating that 3-MA has no effect on radiosensitivity in either H460wt or H460crp53 cells. Cells were pretreated with 3-MA for 3 h followed by radiation exposure. Panel D: Influence of 3-MA on radiation sensitivity. Clonogenic survival assay indicating that 3-MA has no effect on radiosensitivity in either cell line. Cells were pretreated with 3-MA for 3 h followed by an additional 11 days postirradiation. Panel E: Influence of 3-MA on radiation-induced apoptosis. Annexin V/PI staining showing that 3-MA has no effect on radiation-induced apoptosis in either cell line. Cells were pretreated with 3-MA for 3 h prior to irradiation and apoptosis was assessed after 2 days. Unless stated otherwise, data were from three independent experiments. *P < 0.05, irradiated only cells vs. irradiated and 3-MA-treated cells.

FIG. 4. — 3-MA fails to alter radiation sensitivity in H460wt or H460crp53 cells. Panel A: Inhibition of autophagy by 3-MA. Western blot showing autophagy blockade by 3-MA (1 mM) based on levels of LC3 II. Cells were pretreated with 3-MA for 3 h prior to irradiation and protein was collected 3 days postirradiation. The bar graph in each panel indicates the relative band intensity generated from densitometric scans of two independent experiments in arbitrary densitometric units. Panel B: Inhibition of autophagy by 3-MA. Cells were pretreated with 3-MA for 3 h prior to irradiation and cells were stained with acridine orange 3 days postirradiation (scale bar = 200 μm). Panel C: Influence of 3-MA on radiation sensitivity. Cell viability assay indicating that 3-MA has no effect on radiosensitivity in either H460wt or H460crp53 cells. Cells were pretreated with 3-MA for 3 h followed by radiation exposure. Panel D: Influence of 3-MA on radiation sensitivity. Clonogenic survival assay indicating that 3-MA has no effect on radiosensitivity in either cell line. Cells were pretreated with 3-MA for 3 h followed by an additional 11 days postirradiation. Panel E: Influence of 3-MA on radiation-induced apoptosis. Annexin V/PI staining showing that 3-MA has no effect on radiation-induced apoptosis in either cell line. Cells were pretreated with 3-MA for 3 h prior to irradiation and apoptosis was assessed after 2 days. Unless stated otherwise, data were from three independent experiments. *P < 0.05, irradiated only cells vs. irradiated and 3-MA-treated cells.