Figure 7. VEGFR2-β3 integrin-neuropilin-1 interact in vivo and in cultured podocytes. VEGF knockdown decreases alpha_Vbeta₃ integrin activity.

(A) VEGFR2 - beta₃ integrin - neuropilin1 (NRP1) co-immunoprecipitate in vivo, shown by reciprocal VEGFR2 and beta₃ integrin IP. Negative control is rabbit serum (RS). Immunoprecipitates were blotted with VEGFR2, beta₃ integrin, Y¹¹⁷⁵-VEGFR2, anti S⁷⁸⁵- beta₃ integrin and neuropilin-1 antibodies. (B) VEGFR2 - beta₃ integrin – NRP1 co-immunoprecipitate in cultured podocytes. IPs were performed as described in (A) using VEGF^KD podocyte lysates. (C) Dual-immunostaining shows decreased active alpha_vbeta₃ integrin (WOW-1) and total beta₃ integrin in glomeruli from VEGF knockdown mice. (D) Immunocytochemistry shows decreased active alpha_Vbeta₃ integrin (WOW-1) in VEGF knockdown as compared to control podocytes, blue nuclei (Hoechst 33342). Scale bars = 20 µm.