Additional file 6.

Specificity of knockdown by Plexin-A2 shRNA. An embryo was electroporated at E2 (HH stage 13–15) in the ventral neural tube with an shRNA vector targeting Plexin-A2. The effect at HH stage 23 of shRNA electroporation (a) on expression of (b) Plexin-A1, (c) -A2 and (d) -A4 was assessed on adjacent transverse cryosections of hindlimb region spinal cord (30 μm). Plexin-A2 shRNA was effective in reducing Plexin-A2 mRNA (c) (marked knockdown evident in regions enclosed by dotted lines in (a,c)) but was without effect on expression of Plexin-A1 (b) and Plexin-A4 (d). (e) To more precisely map the region of Plexin-A2 shRNA expression onto the Plexin-A2 gene expression pattern, EGFP was visualised by GFP antibody labelling (green) post hybridisation on a section from the same embryo as above. The fluorescent antibody signal was superimposed on a false colour transformation of the Plexin-A2 in situ signal ((e,f) red). This shows that when superimposed (e) there is little overlap of the two colours, confirming the efficacy of the Plexin-A2 knockdown. Bar = 100 μm.

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Bron et al. Neural Development 2007 2:21   doi:10.1186/1749-8104-2-21