Fig. 5

Twist1 knockdown suppressed NE-induced mesenchymal-like phenotype and migration of glioma cells. Lentiviruses were used to establish the stable glioma cell lines with shTwist1. The levels of Twist1, mesenchymal markers and the migration ability were detected in the indicated cells. A-B Real-time PCR (left panel) and western blot (right panel) were used to detect Twist1 expression. The mRNA and protein level of Twist1 significantly decreased in U251 (A, n = 3, p = 0.036) and LN229 cell lines (B, n = 3, p = 0.005). Student’s t-test was used for statistical analysis. C-D NE-induced upregulation of mesenchymal markers was blocked by shTwist1 lentivirus transfection both in U251 (C, n = 3, N-cadherin p = 0.014, Fibronectin p = 0.016, Vimentin p = 0.021) and LN229 cells (D, n = 3, N-cadherin p = 0.037, Fibronectin p = 0.011, Vimentin p = 0.008). One-way ANOVA analysis was used for statistical analysis. E-F Twist1 knockdown offset the pro-migration effects of NE in U251 (n = 5, p = 0.0001) and LN229 cells (n = 5, p = 0.001). One-way ANOVA analysis was used for statistical analysis. Scale bar = 50 μm, *p < 0.05, **p < 0.01