Fig. 4: FOXP4 functions as a positive regulator of the Wnt/β-catenin signaling pathway in OV.

A The RNA-seq data of 426 OV samples from TCGA was subjected to GSEA, the HALLMARK_WNT_BETA_CATENIN_SIGNALING was enriched in high FOXP4 expression group. B Differentially expressed genes in WT and FOXP4-KO cell lines were analyzed, and volcano plots are expressed as up-regulated and down-regulated genes with |LogFC| >1.2 and P < 0.05. C The table indicates that 871 genes are up-regulated and 385 genes are down-regulated. D KEGG analysis of the cells in FOXP4-KO revealed significant changes in the Wnt pathway. E GSEA plot of enriched pathways in RNA-seq data from FOXP4-KO and WT. F GSEA plot of enriched pathways in RNA-seq data of 426 OV samples from TCGA. G The luciferase activities of TCF/LEF1 in A2780 and OVCAR8 cells transfected with pLKO or FOXP4-shRNAs were measured, n = 3. ***p < 0.001 vs pLKO. H The luciferase activities of TCF/LEF1 in A2780 and OVCAR8 cells transfected with EV or FOXP4 were measured, n = 3. **p < 0.01 vs pLKO, ***p < 0.001 vs pLKO. I Real-time PCR was used to measure mRNA levels of FOXP4, CCND1, GSK3B, and c-Myc in A2780 cells stably expressing either control vector (pLKO) or shRNAs targeting FOXP4, n = 3. **p < 0.01 vs pLKO, ***p < 0.001 vs pLKO. J Immunoblot analysis was performed to evaluate the protein levels of FOXP4, CCND1, GSK3B, and c-Myc in cells stably expressing pLKO or shRNAs targeting FOXP4. K Real-time PCR was used to determine the expression of FOXP4 and Wnt/β-catenin target genes in A2780 cells transfected with EV or FOXP4 with/without ICG-001 treatment for 24 h, n = 3. ***p < 0.001 vs pLKO. L Immunoblot analysis was used to detect the protein levels of FOXP4, CCND1, GSK3B, and c-Myc in A2780 cells with stable expression of EV and FOXP4 or overexpression of FOXP4 or ICG-001. M Cell viability determination was performed on the cells stably expressing EV and FOXP4 after treatment with DMSO or ICG-001, n = 3. ***p < 0.001. N The Colony formation numbers of cells in (M) in colony experiments, n = 3. ***p < 0.001. O Relative invasion rates of cells in (M) in a transwell assay, n = 3. ***p < 0.001.