Figure 5. IL-33 interacts with ST2 to activate JNK-enhanced invasion, EMT and stemness. (A) IL-33 and ST2 interaction network from string. (B) Effects of IL-33 on ST2 expression and anti-ST2 blocked the IL-33-induced JNK activation by western blot. (C) Effects of anti-ST2 on the role of IL-33-induced glioma invasion. Glioma cell lines were subject for transwell assay. (D) IL-33 (20ng/mL) and/or anti-ST2 antibody (1 ug/mL) were added in the cell culture. Results are expressed as the mean ± SD; n=3; ***, P < 0.001. (E) Effects of anti-ST2 on the role of IL-33-stimulated EMT related protein expression. Glioma cells were cultured with IL-33 (20 ng/mL) and/or anti-ST2 antibody (1 ug/mL) for 48 hours. The expression of N-cadherin, E-cadherin, Vimentin and β-catenin were quantified by western blot. (F) Effects of anti-ST2 on the role of IL-33-induced glioma stemness. Anti-ST2 blocked the IL-33-induced sphere formation. Representative images of spheres of glioma cells are shown. Scale bar, 100 μm. (G–H), The mean numbers and diameters of spheres were counted and analyzed. Results are expressed as the mean ± SD; n=3; **, P < 0.01; ***, P < 0.001. (I) The levels of stemness related genes including CD133, Nestin and Oct4 were detected by Western blot. Results are shown as mean of the data from at least three independent experiments.