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. 2021 Feb 15;131(4):e139542. doi: 10.1172/JCI139542

Figure 1. IDH1-R132H inhibition in mIDH1 mouse-NSs and human-GCs confers radiosensitivity and promotes release of DAMPs.

Figure 1

(A) Mouse-NS cultures were generated from WT-IDH or mIDH1 glioma models. (B–E) Clonogenic assay of mIDH1 and WT-IDH mouse-NSs and human-GCs. (B and C) Mouse-NS were treated with 0–8 Gy ionizing radiation (IR) and 1.5 μM AGI-5198 (red) or DMSO vehicle (blue). (D and E) Human-GCs SJGBM2 (WT-IDH) and MGG119 (mIDH1) were treated with 0–20 Gy IR and 5 μM AGI-5198 (red) or DMSO vehicle (blue). ****P < 0.0001, nonlinear regression. Bars represent mean ± SEM (n = 3 technical replicates). Representative images of colonies stained with crystal violet in each treatment group are shown under the survival fraction graphs. (F–I) Calreticulin (CRT), ATP, and HMGB1 expression levels within mIDH1 mouse-NSs and human-GCs. Mouse-NS were treated with 3 Gy IR and 1.5 μM AGI-5198 for 72 hours. Human-GC were treated with 10 Gy IR and 5 μM AGI-5198 for 72 hours. Quantification of CRT expression on mIDH1 mouse-NSs and human-GCs after treatment is shown in F and G, respectively. Representative histograms display CRT marker’s expression levels (black, isotype control; green, DMSO vehicle; light blue, IR; red, AGI-5198; dark blue, AGI5198 + IR). ****P < 0.0001, **P < 0.001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates). (H) Quantification of ATP release in the supernatant of mIDH1 mouse-NSs and human-GCs. *P < 0.01; **P < 0.01; ***P < 0.001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates). (I) Quantification of HMGB1 release in the supernatant of mIDH1 mouse-NSs and human-GCs. **P < 0.01; ***P < 0.001; ****P < 0.0001, 1-way ANOVA. Bars represent mean ± SEM (n = 3 technical replicates).