Fig. 6.

Antitumor effect by combined inhibition of EFTUD1 with an autophagy blocker. (A) Effect of combined treatment (EFTUD1 siRNAs and CQ) on glioma cell growth. CQ (50 μM) was added to siRNA-transfected cells 24 hours before analysis. Cell viability analysis was performed 0, 2, 4, and 6 days after transfection. Results are presented as fold relative to the cell viability in control siRNA-treated cells at day 0. Data are mean ± S.D. (bars) of 3 experiments. **P < 0.01 using 1-way ANOVA. (B) Effect of combined treatment on clonogenicity in U87MG glioma cells. Twenty-four hours after transfection, cells were treated with CQ (50 μM) for 3 hours (combined treatment) and then plated out to grow on soft agar in 6-well plates at 2 × 10⁴ cells/well. Colonies stained with MTT were counted 4 weeks after siRNA transfection. Results are mean ± S.D. (bars) of 3 experiments. n.s. not significant, *P < .05, **P < .01 using 1-way ANOVA. (C) Effect of combined treatment on apoptosis in U87MG glioma cells. CQ (50 μM) was added 24 hours before analysis (combined treatment). Caspase 3/7 activity in U87MG glioma cells was analyzed 6 days after siRNA transfection. Caspase 3/7 activity was normalized to viable cell signals measured by the CellTiter Glo luminescent cell-viability assay. Relative caspase-3/7 activity is presented as fold relative to this normalized value found in control siRNA-transfected cells. Results are mean ± S.D. (bars) of 3 experiments. n.s. not significant, *P < .05, **P < .01 using 1-way ANOVA.