Fig. 8

Effect of RBP4 on sprouting in vivo and in vitro. HUVECs were treated with 10 ng/ml RBP4 for 48 h, with or without 1 μM Ki8751 pretreatment for 12 h. A Relative mRNA expression of ANGPT2, TIE1, HEY1 and DLL4 in RBP4-treated HUVECs (n = 3). B Relative mRNA expression of ANGPT2, TIE1, HEY1 and DLL4 in the control, RBP4 and RBP4 + Ki8751 groups (n = 3). C, D Protein levels of VEGFR2 in the three groups (n = 3). E Immunofluorescence staining of DAPI (blue) and VEGFR2 (red) in HUVECs. Scale bar: 50 μm. F Co-IP examination of VEGFR2-RBP4 interaction. H, J, K Retinal IB4 staining of P14 in OIR and relative number of tip cells as well as filopodia. Scale bar: 200 μm (n = 3). G, I Choroidal sprouting analysis in three groups (n = 3). Scale bar: 500 μm. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001