Fig. 2

CircRYK induces GBM cell epithelial-mesenchymal transition and GSC maintenance in vitro. (A) qRT‒PCR was utilized to examine the levels of circRYK expression in GBM cells and NHAs. (B) At splice junctions, designated shRNAs for circRYK are illustrated schematically. (C-D) qRT‒PCR was used to evaluate the expression of circRYK in GBM cells transfected with circRYK-targeting shRNA. (E) Transwell assays for cell EMT research following glioma cell transfection with expression vectors or shRNA. (F) The invasion of U87-GFP and pGBM-1-GFP tumor spheres was observed using three-dimensional spheroid experiments. The quantitative findings are shown together with representative photographs taken three days after plating. (G) The migration of GBM cells was examined via a wound healing assay. (H) After transfection, the expression of N-cadherin, Vimentin, Snail, and MMP-9 in GBM cells was analysed via Western blotting. (I) The stemness of GSCs following transfection was evaluated via a clonogenic trial. (J) The CellTiter-Glo assay was used to assess the effects on GBM-GSC cells. (K) Western blots were performed to assess the expression of Sox2, Oct4, and Nanog in GBM-GSC cells after transfection. (L) Immunofluorescence staining was applied to determine the expression of N-cadherin and Vimentin in treated U87 cells. Scale bar, 100 μm. Each experiment was performed three times, and the results are displayed as the mean ± SD. CONT, control (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)