Fig. 3. GLT8D1 is important for glioma stem cell maintenance.
a, b Indicated cells were stained with a PE-labeled anti-AC133 (130-113-186, Miltenyi Biotec) and FITC-labeled anti-CD44 (75122 S, CST) antibody followed by flow cytometry analysis (n = 3). b Quantification data for a. c Knockdown of GLT8D1 inhibits GSC marker gene expression in GSC11 cells verified by IB. Indicated antibodies were used. d–f Representative images of GSC11 and GBM1 tumor sphere expressing GLT8D1 shRNAs or control shRNA are shown (d). e, f Quantification data for d. Scale bar: 500 μm. g, h A subcutaneous limiting dilution tumor formation assay (1 × 102, 1 × 103, 1 × 104 and 1 × 105 cells/per mouse) was performed in indicated cells, tumor masses were harvested after 11 weeks. h Quantification data of tumor incidence for g. i Survival analysis of mice intracranially implanted with 5 × 105 indicated cells. X-axis represents days after cell injection. The survival difference was evaluated by log-rank analysis. n = 7. j Representative images of H&E staining for tumor formation of indicated xenografts for i. Scale bar: 2 mm. T = Tumor tissues, N = Normal tissues. Means ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, t-test.