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Fig. 7. Distribution of neuropilin 2 ligands in normal and in Sema3F- or neuropilin 2-transfected embryos. (A-F) Double staining with NP2dC-AP (A,B,E; dark blue) and anti-neurofilament antibody (C,F; magenta) on normal embryos at stage 17(A-C) and stage 19 (D-F). (D) Images in E and F merged. Stained brain specimens were flat-mounted and focused on the MHB (B-F). Trochlear nucleus along the rostrocaudal axis is indicated by double-headed arrows in C,F. Trochlear axons extend in the isthmic region devoid of NP2dC-AP staining. (G-J) Detection of GFP (G,I; green) and NP2dC-AP staining (H,J; dark blue) on Sema3F-(G,H) or neuropilin 2-(I,J) transfected embryos. NP2dC-AP can detect misexpressed Sema3F (G,H). NP2dC-AP staining was dismissed in the brain where neuropilin 2 was misexpressed (I,J). NF, neurofilament. Scale bars: 1 mm in A,G-J; 200 µm in B-F.





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