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. 2017 Mar 20;127(4):1425–1437. doi: 10.1172/JCI90644

Figure 4. Decreased chemokines and chemotaxis of T cells to IDH-MUT–derived CM compared with media derived from IDH-WT cells.

Figure 4

(A and B) GL261-WT or GL261- MUT cells treated with either DMSO (control) or 1 μM IDH-C35 for 3 days were stimulated with 10 ng/ml recombinant murine IFN-γ for 6 hours prior to analysis. (A) RT-PCR analysis of Cxcl9 and Cxcl10 from GL261-WT (WT) and GL261-MUT (MUT) cells treated with or without IDH-C35. Gene expression levels were normalized to Gapdh and are shown relative to levels in WT samples. (B) Chemotaxis of activated CD8+ T cells toward GL261 glioma cell–CM by Transwell assay. CM were diluted 1:1 with fresh media. For CXCL10 conditions, 100 ng/ml recombinant murine CXCL10 (rCXCL10) was added to GL261-MUT–CM. For blockade of CXCR3, T cells were preincubated in 10 μg/ml anti-CXCR3 for 30 minutes prior to use. Results are representative of 3 independent experiments, each performed in triplicate with highly similar results. (C) RT-PCR analysis of OAS2 and CXCL10 gene expression from 4 unique glioma patient–derived neurospheres with IDH1 mutations either untreated (–) or treated with IDH-C35 for 3 days. Each line (and sample name) corresponds to a neurosphere generated from a different patient. Gene expression levels were normalized to GAPDH and are shown relative to expression levels in untreated neurosphere NCH1681. P values were obtained using 1-way ANOVA with a Newman-Keuls multiple comparisons test (A and B) and a 2-tailed, paired t test (C). *P < 0.05, **P < 0.01, ***P < 0.001; NS, P > 0.05.