Fig. 6. Binding of C. elegans semaphorins to PLX-1 expressed in the cultured cells. (A) A western blot of PLX-1 expressed in HEK293T cells. An immunoreactive band the size of 220 kDa was detected for Myc-PLX-1 with anti-Myc antibody. The predicted size of the peptide is 210 kDa. (B) A western blot of C. elegans semaphorins secreted in the culture medium of HEK293T cells. An immunoreactive band the size of 143 kDa (lane 1) and 110 kDa (lane 2) for Ce-Sema-1a-C-Fc-AP and Ce-Sema-2a-Fc was detected, respectively, with anti-Fc antibody. The predicted size of each peptide is 139 kDa and 94 kDa, respectively. (C) Scatchard analysis of the binding of Ce-Sema-1a-C-Fc-AP to PLX-1. The inset shows the binding curves of Ce-Sema-1a-C-Fc-AP to PLX-1 expressed on HEK293T cells (the upper line) and to HEK293T cells transfected with pCAGGS as a control (the lower broken line). (D) Binding of Ce-Sema-1a-C-Fc-AP to PLX-1. (i,ii,iii) Untransfected HEK293T cells. (iv,v,vi) HEK293T cells expressing PLX-1. (i,iv) Cells were reacted with the anti-Myc antibody. (ii,v) Cells were reacted with Ce-Sema-1a-C-Fc-AP. (iii,vi) Cells were reacted with Ce-Sema-2a-Fc. Scale bar: 100 µm.