Figure 5 | Scientific Reports

Figure 5

From: Interplay between CCN1 and Wnt5a in endothelial cells and pericytes determines the angiogenic outcome in a model of ischemic retinopathy

Figure 5

Adenovirus-mediated expression of CCN1 induced Wnt5a expression and activity in cultured pericytes. (A) Cultured ECs and pericytes were transduced with an adenoviral vector expressing either luciferase (Ad-luc) or CCN1 (Ad-CCN1). After incubation in serum-free medium for 16 h, cell lysates were analyzed by western immunoblotting for CCN1 protein expression. (B,C) Expression Wnt ligands was analyzed by qPCR in Ad-luc- and Ad-CCN1-infected cells. *p < 0.001 versus Ad-luc. (D,E) The relative abundance of Wnt5a transcripts and Wnt3a and Wnt7a was calculated by comparing their ΔCTs in pericytes versus ECs. (F,G) Effects of CCN1 on canonical and non-canonical Wnt signaling activity. Following adenoviral-mediated gene transfer with either Ad-CCN1 or Ad-luc, cultured pericytes were transiently transfected with either pGL3-NFAT luciferase reporter or LEF/TCF-M50 Super 8× TOP Flash plasmid. Luciferase activity was determined in cell lysates and media. Values shown are from a representative experiment performed in triplicate. **p < 0.01 versus Ad-luc. Experiments were repeated three times using different cell preparations with similar results.

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