Figure 2.

Effect of NSC-sTRAIL in combination with Lan C on GBM cells and stem-like cells. (A): 3 × 10³ NSCs, U87, Gli36, GBM4, GBM6, or GBM8 GSCs expressing Gluc were treated with 0–3,000 nM of Lan C. Cell viability was monitored 3 days later by assaying Gluc activity in the medium. Data presented as the mean Gluc RLU/ml ± SD (n = 8; *, p <.05; **, p <.01 vs. no treatment by two-way ANOVA). (B, C): 3 × 10³ U87-Guc cells (B) or Gli36-Gluc cells (C) were cocultured with 300 NSC-sTRAIL cells and treated with Lan C (0.25 μM) or DMSO. GBM cell viability was monitored using Gluc assay 48 hours later (**, p <.01 vs. control; ^##, p <.01 vs. Lan C alone using ANOVA and Tukey’s post hoc test). (D): 3 × 10³ GBM4, GBM6, or GBM8 GSCs (~60 small neurospheres), or HA expressing Gluc were seeded were cocultured with 300 NSC-sTRAIL cells and/or 0.1 μM Lan C. Cell viability was monitored by measuring Gluc activity in the medium 48 hours post-treatment. Data presented as the mean relative Gluc RLU ± SD (n = 8) in which cells cocultured with NSC-Vluc control is set at 100 (*, p <.05 and **, p <.01 vs. control; ^##, p <.01 vs. Lan C alone using ANOVA and Tukey’s post hoc test). (E): 3 × 10³ GBM8-Gluc cells (~60 small neurospheres) were seeded in lower well of a 96-well 0.4 μm Transwell plate in NBM E/F20. Different amounts (3, 30, 300, or 3,000) of NSC-sTRAIL cells were seeded in the upper wells. After 12 hours, GBM cells were treated with DMSO (control), 0.1, or 1 μM of Lan C and viability was monitored 48 hours later using the Gluc assay. Data presented as the mean Gluc RLU/ml ± SD (n = 8; **, p <.01 vs. no Lan C; ^#, p <.05; ^##, p <.01 vs. no coculture with NSC cells). Abbreviations: DMSO, dimethyl sulfoxide; GBM, glioblastoma; HA, human astrocytes; NSC, neural stem cell; TRAIL, tumor necrosis factor-related apoptosis-inducing ligand.