Fig. 6

The effects of FH535 (a canonical Wnt signaling inhibitor) and Foxy-5 (a Wnt5a agonist) on Wnt signaling pathway and EMT in ARPE-19 cells. A Western blot analysis of fibronectin, Wnt5a, active β-catenin, Naked1, Dvl2, and Wnt3a in ARPE-19 cells treated with TGFβ1 (10 ng/mL) alone or in combination with FH535 (0.5 μmol/L) or Foxy-5 (50 and 100 μmol/L), as well as in untreated ARPE-19 cells with or without Foxy-5 (50, 100, or 200 μmol/L). B–F Quantitative analysis of Western blot analysis of fibronectin, Wnt5a, active β-catenin, Naked1, and Dvl2 was performed in indicated groups with GAPDH served as loading control. G–I Expression and quantitative analysis of active β-catenin protein in the nuclear and cytoplasmic extracts of ARPE-19 cells under different conditions; Lamin B1 and GAPDH were used as nuclear- or cytoplasmic-specific protein loading controls, respectively. J–K The representative images of immunofluorescence for fibrotic marker fibronectin (red) or active β-catenin (green), with DAPI (blue) labeling all nuclei, were obtained from Vehicle (0.005% DMSO), TGFβ1 + Vehicle (0.005% DMSO), TGFβ1 + FH535 (0.5 μmol/L), Vehicle + Foxy-5 (200 μmol/L) groups. Scale bars = 75 μm in (J) and Scale bars = 50 μm in (K). Dvl, dishevelled; NC, negative control; TGFβ1, transforming growth factor beta 1. All results performed above are presented as mean ± SE from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared with TGFβ1-treated ARPE-19 cells; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 compared with untreated ARPE-19 cells