Testing for transgene silencing. (A-B″) Live confocal images of 5 dpf larvae carrying two transgenes ins:hmgb1-eGFP (labels β-cell nuclei green) and ins:mCherry NTR2.0 (designed for β-cell expression of NTR2.0 and mCherry). Both transgenes mark islet cells in the head of the pancreas (20×, scale bar: 100 µm) (A). Close up of green nuclei (B) and red cytoplasm (B′) and co-expression in β cells (B″), indicating correct β-cell-specific expression of NTR2.0 (40×, scale bar: 10 µm). (C-C″) Confocal images of sections from ins:mCherry NTR2.0 fish with immunofluorescent detection of insulin (to detect β cells, green; C) and mCherry (red, reports on NTR2.0 co-expression; C′); complete colocalization can be seen throughout the islet (C″) (40×, scale bar: 10 µm). a, anterior; d, dorsal; p, posterior; v, ventral.
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