Fig. 5: PTK7 is a transcriptional target of FOXP4 in OV.

A The mRNA correlation between FOXP4 and PTK7 in OV from TCGA database. B The spearman correlation between the expression of FOXP4 and PTK7 in all tumor types in TCGA. Red represents positive expression and green represents negative expression. C Real-time PCR analysis of FOXP4 and PTK7 mRNA levels in A2780 cells transfected with EV or FOXP4 expression vector, n = 3. **p < 0.01 vs EV, ***p < 0.001 vs EV. D Immunoblot analysis showing FOXP4 and PTK7 protein expression in A2780 cells transfected with EV or FOXP4. E Immunoblot analysis of FOXP4 in A2780 and OVCAR8 cells stably expressing either pLKO or shRNAs targeting FOXP4. F Schematic diam shows human PTK7 gene promoter and a putative FOXP4 binding site. TSS: transcription start site. G Schematic diam shows the human PTK7 WT and Mut promoters. The potential binding sites for FOXP4 were highlighted with yellow. H The activities of the human PTK7 WT and Mut promoters in 293T cells transfected with EV or FOXP4 was measured, n = 3. ***p < 0.001 vs EV. I Two shFOXP4s were co-transfected with the indicated plasmid into 293T cells for 36 h. The luciferase activity was then measured, n = 3. ***p < 0.001 vs pLKO. J, K Chip-PCR assay detecting FOXP4 binding to the human PTK7 promoter in A2780 cells expressing stable FOXP4, n = 3. ***p < 0.001 vs IgG. L A2780 cells were treated with ICG-001 or LiCl to detect PTK7 protein levels. M The luciferase activities of TCF/LEF1 in A2780 and OVCAR8 cells transfected with pLKO or shRNAs-PTK7 were measured, n = 3. **p < 0.01 vs pLKO, ***p < 0.001 vs pLKO. N The luciferase activities of TCF/LEF1 in A2780 and OVCAR8 cells transfected with EV or PTK7 were measured, n = 3. **p < 0.01 vs EV.