Figure 12

Co-immunoprecipitation (IP) experiments showing homophilic interaction of human and murine B3 and no interaction of B3 with plexins A1, B1 or B2. Total lysates and IPs were analyzed by Western blot (WB) using antibodies as indicated in the figures. IP was performed with pAbB3-B against human B3 and shows interaction between mouse and human B3 and no interaction between B3 and human plexins A1, B1 and B2. (A) Cells were co-transfected with pSecTag2B/B3 encoding myc-tagged human full-length B3 and pcDNA3.1/mB3 encoding V5-tagged mouse B3 lacking most of its intracellular part. Cells transfected with pcDNA3.1/mB3 and pSecTag2B vector without insert served as negative control. (B) COS-7 cells were co-transfected with pIRES/B3 encoding non-tagged full-length human B3 and pcDNAVSV/A1 encoding VSV-tagged full-length human plexin A1. Cells co-transfected with pcDNAVSV/A1 and pIRES vector without insert served as negative control. (C) COS-7 cells were co-transfected with pIRES/B3 and pcDNAVSV/B1 encoding VSV-tagged full-length human plexin B1. Cells co-transfected with pcDNAVSV/B1 and pIRES vector without insert served as negative control. (D) COS-7 cells were co-transfected with pIRES/B3 and pFLAG/B2 encoding FLAG-tagged full-length human plexin B2. Cells co-transfected with pFLAG/B2 and pIRES vector without insert served as negative control.