Figure 5.

HDGF mediates ZEB1 to promote EC migration and invasion by modulating β-catenin-induced EMT signal. A and B. Strong interaction between HDGF and β-catenin, TCF4, showing by reciprocal CoIP through using antibody (anti-TCF4, anti-β-catenin) respectively. C. EC cells were immunostained with anti-HDGF, TCF4, β-catenin and visualized under fluorescence microscope. Scale bar: 20 µm. D. CoIP assay showed that less TCF4 bound to β-catenin after suppression of HDGF compared to control group. Similarly, decreased HDGF protein induced less β-catenin binding to TCF4 by CoIP assay. β-actin served as internal control. E and F. Suppress HDGF decreased nuclear translocation of β-catenin, while HDGF over-expression had the opposite effect. β-actin served as the cyto internal control. Lamin B1 was considered as nuclear loading control. G. HDGF over expression enhanced whereas HDGF silencing suppressed TOP flash reporter activity, respectively. H. Western blotting analysis for the expression levels of HDGF, β-catenin, TCF4, ZEB1, and EMT-associated proteins in EC cells with different treatments. Data were shown as the mean ± SD, **P < 0.01; ***P < 0.001.