Endogenous chymotrypsinogen B1in β-cells. (A) Schematic of experimental design for generation of Tg(CTRB1-GFP),Tg(insa:mCherry) using CRISPR/Cas9 and homology directed repair for insertion of GFP at C-terminus of endogenous ctrb1 gene. (B) Brightfield image of Tg(CTRB1-GFP),Tg(insa:mCherry) animal at 5 dpf. (C) GFP fluorescence in Tg(CTRB1-GFP), Tg(insa:mCherry) animal at 5 dpf indicating successful fluorescence in Tg(CTRB1-GFP),Tg(insa:mCherry) animal at 5 dpf indicating successful fluorescence in Tg(CTRB1-GFP),Tg(insa:mCherry) animal at 5 dpf. (E) Quantification of colocalization of CTRB1 in zebrafish β-cells at 5 dpf by fold change in proportion of GFP+ mCherry+ cells in Tg(Ctrb1-GFP),Tg(insa:mCherry) compared with Tg(ptf1a:GFP),Tg(insa:mCherry). Error bars represent s.e.m. ***P<0.001, Student's t-test.
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