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Fig. 8 | Molecular Cancer

Fig. 8

From: A novel protein AXIN1-295aa encoded by circAXIN1 activates the Wnt/β-catenin signaling pathway to promote gastric cancer progression

Fig. 8

CircAXIN1 activates the Wnt/β-catenin signaling pathway. a-b CircAXIN1 and AXIN1-295aa activate Wnt/β-catenin-dependent transcription. a Topflash/Fopflash assays were performed in 293 T and AGS cells with circAXIN1 transfection. b Topflash/Fopflash assays were performed in 293 T and AGS cells with linear-295aa transfection. c Topflash/Fopflash assays were performed in 293 T cells with individual circAXIN1 siRNA transfection. d Inhibition of circAXIN1 causes downregulation of β-catenin in both the cytoplasm and nucleus. Expression of β-catenin in the cytoplasm and nucleus with individual circAXIN1 siRNA transfection. e CircAXIN1 overexpression enhances β-catenin nuclear translocation and circAXIN1 siRNA inhibits β-catenin nuclear translocation. Immunofluorescence localization of β-catenin was detected with circAXIN1 or circAXIN1 siRNA transfection. Wnt agonist1 and XAV939 (a Wnt/β-catenin signaling inhibitor) served as positive controls. f CircAXIN1 overexpression activates the downstream gene expression of the Wnt/β-catenin pathway. g Individual circAXIN1 siRNA transfection leads to the reduced expression of Wnt/β-catenin pathway downstream genes. h The mRNA levels of c-Myc, c-Jun, CD44, Met, cyclinD1, and VEGF decrease with each circAXIN1 siRNA transfection. i More β-catenin binds to the TCF-dependent promoter of Wnt/β-catenin downstream genes with overexpression of circAXIN1. A ChIP assay was performed using anti-β-catenin to detect the binding of β-catenin to the promoters of c-Jun, c-Myc, and CD44. C-Jun1/2 means region1 or 2 of the c-Jun promoter. j Linear AXIN1-295aa rescues the inhibitory effect of circAXIN1 siRNA on Topflash activity. Topflash/Fopflash assays were performed with circAXIN1 siRNA transfection or co-transfection of circAXIN1 and linear-295aa. k Linear AXIN1-295aa elevates the downregulation of genes downstream of the Wnt/β-catenin pathway caused by circAXIN1 siRNA transfection

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