Fig. 5. NRP2 is predominantly required for SEMA3F function in LEC.

A) Western blot of endogenous NRP1 and NRP2 in LEC in the presence of control (siCtrl), NRP1, or NRP2 siRNA. B) Fluorescent imaging of LEC transfected with the indicated siRNA and then treated with vehicle or 100 ng/ml SEMA3F. Parental and siCtrl-transfected LEC treated with increasing doses of SEMA3F and evaluated for collapse (C), or attachment (D) For both collapse and attachment assays, values are reported relative to vehicle control and quantification was done by ImageJ, based on 25 imaging fields each from quadruplicate wells in three independent experiments. E) IC₅₀ of SEMA3F was calculated from the dose curve generated in (C) and (D). Statistical significance was determined using one-way ANOVA, *p<0.05, **p<0.01, ***p<0.001.