Knockdown of plxna3 results in aberrant motor neuron patterns.

The motor neurons of 24 hour post fertilization sidetracked zebrafish embryos (top) exhibit ectopic motor neuron exit points from the motor cord (arrows) compared to WT embryos (bottom). Asterisks indicate endogenous motor neuron exit points. Embryos are oriented anterior (left) to posterior (right).

(A) Anti-GFP (1:1000 dilution, Clontech) western blot confirming expression of PlxnA3-GFP² in the BRET² assay. The expected molecular weights are 176 kDa and 27 kDa for PlxnA3-GFP² and GFP², respectively. (B) Anti-RLuc (1:2500 dilution, Millipore) western blot confirming expression of PlxnA3-RLuc in the BRET² assay. The expected molecular weights are 185 kDa and 36 kDa for PlxnA3-RLuc and RLuc, respectively. (C) Anti-FLAG staining of COS-7 cells expressing FLAG-tagged Nrp2a. No fluorescence was observed in mock-transfected cells. The scale bar in the bottom left frame is the same for all images. (D) Dot blot confirming alkaline phosphatase activity in media from cells transfected with alkaline phosphatase-tagged SEMA3F. (E) Confirmation of alkaline-phosphatase-tagged SEMA3F binding to COS-7 cells expressing Nrp2a and PlxnA3.