Fig 2: Overexpression of CUX1 enhanced the activity of Wnt/ß catenin signaling pathway in glioma cells. (A–C) Gene set enrichment analysis (GSEA) pathways enriched by CUX1. (A) The dotplot of partially enriched pathway. (B) The dotplot of the up- and down regulated pathways. (C) The up-regulated pathway of Wnt/ß-catenin. (D) Wnt/ß-catenin pathway was enriched and upregulated in glioma by gene set variation analysis (GSVA) analysis. (E,F) Immunoflurescence showed that overexpression of CUX1 increased expression of Axin2 proteins in cytoplasm of TJ905 cells compared to empty group. Meanwhile, CUX1 overexpression also increased the expression of ß-catenin in nucleus and cytoplasm of TJ905 cells compared to control.

Fig 3: Inhibiting Axin2 or ß-catenin could partially reverse the malignant phenotype caused by CUX1 overexpression in gliomagenesis. (A,B) Western blot analysis presented the levels of subunits in Wnt/ß-catenin signaling pathway after knocking-down Axin2 or ß-catenin in TJ905 cells ectopically overexpressing CUX1. (C) CCK8 assay was used to detect the change of proliferation capability after inhibiting Wnt/ß-catenin in TJ905 cell lines. *, compared with the empty vector group; #, compared with the empty vector group; a, compared with the empty vector group; ?, compared with the empty vector group. (D,E) Transwell assay to evaluate the reversal effect of silencing Wnt/ß-catenin pathway on the migration activity in TJ905 cell lines. (F,G) The change of proliferation capability after inhibiting Wnt/ß-catenin was detected by colony formation assays in TJ905 cell lines. (H,I) Western blot analysis presented the levels of EMT markers (N-cadherin, E-cadherin, Vimentin and Snail) after inhibiting Wnt/ß-catenin pathway (knocking-down Axin2 or ß-catenin) in TJ905 cells ectopically overexpressing CUX1.