Fig. 1. FGL2 promotes stem-like properties of glioma cells in vivo, but not in vitro.

FGL2^KO, FGL2^low, FGL2^mid, and FGL2^hi GL261 cells were generated for following assays. (A), Expression of FGL2 and stemness-associated transcription factors were detected by Western blotting. Top: Representative blots. Bottom: Quantitative analysis of expression. (B), Expression levels of GSC markers on GL261 cells detected by FACS. (C), In vitro limiting dilution sphere-forming assay. (D), In vitro cell proliferation assay. Data are presented as the mean ± SD (n = 3). (E), Tumor incidence of GL261 cells in WT mice. Cells were orthotopically implanted into the brains of C57BL/6 mice at the indicated numbers. (F), Limiting dilution tumorigenesis assay of GL261 cells in C57BL/6 mice. **P < 0.01, ***P < 0.001, likelihood ratio test. (G-J), Survival curves of C57BL/6 mice (n = 6) implanted with FGL2^KO (G), FGL2^low (H), FGL2^mid (I), or FGL2^hi (J) cells at indicated numbers. (K and L), Survival curves of NSG mice (K) and Scid mice (L) implanted with 2000 FGL2^KO or FGL2^Hi cells (n = 6). All data are representative of at least 2 independent experiments. The survival curves were generated by Kaplan–Meier analysis, and the log-rank test was used to compare overall survival between groups.