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. 2015 Sep 3;6(31):31151–31163. doi: 10.18632/oncotarget.5150

Figure 5. Inhibition of non-canonical glutamine metabolism sensitizes PCSCs to radiotherapy via intracellular ROS accumulation.

Figure 5

A. PANC-1 (left panel) and SW1990 (right panel) spheres were obtained by 14 days of culture, and the culture medium was replaced with fresh medium containing indicated supplements (glutamine, 2.5 mM; dimethyl αKG, 4 mM; OAA, 4 mM) 12 hours before IR treatment. Clone formation was assessed 14 days after irradiation and survival fractions were calculated for each group. B. sphere cells received treatment as described in 5A and the relative DCF fluorescence measured after exposure to 4, 6, or 8 Gy IR. Fluorescence intensity of cells cultured in medium supplemented with 2.5 mM glutamine for each IR group was set as 1. C. shControl, shGLUD1, and shGLS1#2 cells were cultured in sphere-formation condition for two weeks followed by treatment with indicated IR doses. Clone formation was measured 14 days after irradiation. Survival fractions were calculated for each group. D. cells treated as described in 5C were treated with 4, 6, or 8 Gy IR followed by measurement of DCF fluorescence. The fluorescence intensity of shControl group was set as 1. Error bars represent the mean ± S.D. of triplicate experiments. Statistical significance was calculated using the Student's t test or ANOVA. **p < 0.01; ***p < 0.001. ▼, irradiation treatment; ●, timing of evaluation.