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Fig. 2
Anti-zebrafish Gnrh3 Gap polyclonal antibody is specific to zebrafish Gnrh3.
(A) The immunostaining of Gnrh3-ir soma and fibers (white arrow; Aa) in the brain with anti-zebrafish Gnrh3 Gap was eliminated when the antibody was substituted with pre-immune serum (Ab). (B) Immunostaining with anti-Gnrh3 Gap (green; Ba) on brain sections of gnrh3:tdTomato (red; Bb) adults shows co-localization between Gnrh3-ir soma and gnrh3:tdTomato-labeled soma (Bc), indicating the specificity of the antibody to Gnrh3. White arrows indicate Gnrh3-tdTomato-expressing soma that were positively stained by anti-Gnrh3 Gap. (C) Immunostaining with (Ca-Cc) anti-zebrafish Gnrh3 Gap or (Cd) pre-immune serum in COS7 cells transfected with (Ca) control pcDNA3.1 plasmid, (Cb) gnrh2-pcDNA3.1 plasmid, or (Cc,d) gnrh3-pcDNA3.1 plasmid. The cells that express zebrafish Gnrh3 and are immunostained with anti-zebrafish Gnrh3 Gap (red) are indicated by white arrows (Cc). Scale bars = 50 µm.