Figure 7
Inhibition of non-canonical Wnt5a reduced pathological neovascularization following OIR. (A–D) Expression of Wnt5a, Wnt7a, Wnt7b and Wnt3a in mice with either endothelial- or pericyte-specific CCN1 deletion and their wild-type littermates following OIR. Wnt ligand gene expression was analyzed by qPCR in retinal lysates harvested at P17. **p < 0.01 versus CCN1+/+. *p < 0.01 versus Cdh5-CreERT2 CCN1Δ/Δ. (E–G) Flat-mount images of IB4-stained retinas from OIR mice at P17 following treatment with either TNP470 or vehicle alone (Veh). Areas of pre-retinal neovascular tufts at P17 were delineated by a computer-assisted image software and highlighted in blue (c for a and d for b). The compiled data showing the percentage of neovascular and avascular areas are represented in F and G. **p < 0.05 versus +Veh. (H) Transcript levels of CCN1, YAP, SNAIL and G3BP2 as determined by qPCR in retinas from OIR mice injected with either control vehicle or TNP470. RNA levels at P17 (+Veh) were set to 100% to facilitate comparison among groups. *p < 0.01 versus P17 (+Veh). (I) Retinal flat-mounts from OIR CCN1:GFP reporter mice treated with vehicle or TNP470. Note that TNP470 treatment resulted in reduced neovascular tuft formation and reexpression of the CCN1:GFP reporter in ECs instead of perivascular pericytes in remnant tufts (arrows).
