Fig. 2. VLP-NP suppresses Wnt signaling activated by Wnt ligand in cultured cells.
A: HRMEC were transfected with VLN-NP or LN-NP for 48 hr and treated with LCM or WCM for 4 hr (p-LRP6, t-LRP6, n-p-β-catenin, t-β-catenin) or 24 hr (VEGF). Levels of p-LRP6, t-LRP6, n-p-β-catenin, t-β-catenin and VEGF in the cell lysates were measured by Western blot analysis. B–D: Semi-quantification of p-LRP6 and t-LRP6 (B), n-p-β-catenin and t-β-catenin (C), and VEGF (D) levels by densitometry and normalized by β-actin levels. **p<0.01, * p<0.05, n=3. E: HRMEC were transfected with VLN-NP or LN-NP for 48 hr and treated with Mab2F1 (50 µg/ml) 4 hr before adding LCM/WCM for additional 24 hr. VEGF mRNA levels were measured by q-PCR and normalized by 18S. **p<0.01, * p<0.05, NS: not significant, n=3; F, G: HRMEC were transfected with VLN-NP or LN-NP for 48 hr and infected with Ad-GFP/Ad-S37A for additional 24 hr. T-β-catenin and VEGF protein levels were measured by Western blot analysis (F), and VEGF mRNA levels were measured by q-PCR and normalized by 18S (G). H: hTERT-RPE cells were transfected with TOPFLASH vectors and then treated as indicated for 24 hr. TOPFLASH activity was measured using Luciferase assay and expressed as the firefly/renilla ratio relative to that of LCM-LN group. **p<0.01, n=3.
