Fig. 1

Generation of nap1l1 transgenic line in zebrafish. (A) Sequence comparison of upstream intergenic sequences between the zebrafish nap1l1 gene with its human and mouse homologues (https://ecrbrowser.dcode.org). (B) The strategy for the target plasmid pDest-tol2-nap1l1-nEGFP construction with the primer positions of each reconstruction fragment. (C) Amplification of each reconstruction fragment. The DNA templates of lane 1–3 were plasmid pDest-tol2-pA2, zebrafish genomic DNA, and plasmid pME-nEGFP, respectively. Red arrowhead points to the right size of backbone fragment. (D) PCR-based Genotyping confirmed that the fluorescent or fluorescenceless F1 progeny harbor the Tg(nap1l1:nEGFP) transgene. Line1-2, the fluorescenceless F1 progeny of Tg(nap1l1:nEGFP)zs101 and Tg(nap1l1:nEGFP)zs102 respectively; Line3-6, the fluorescent F1 progeny of Tg(nap1l1:nEGFP)zs101, Tg(nap1l1:nEGFP)zs102, Tg(nap1l1:nEGFP)zs103, and Tg(nap1l1:nEGFP)zs104, respectively; Line7-8, each single wildtype embryo; Line9, zebrafish genome was not added as PCR template. (E-G, Ha, Ja-b, Ka-d) Overview of the transgene expression pattern of a zebrafish line Tg(nap1l1:nEGFP)zs102 during embryonic development. (Hb, Ia, Jc-e, Ke-f) In situ hybridization for egfp. (Hc, Ib, Jf-h, Kg-h) In situ hybridization for nap1l1. White arrowhead points to the embryonic shield. Blue or green arrowheads point to the lateral line neuromasts. Abbreviations: ALL, anterior lateral line; h, heart; nm, neuromast; nt, neural tube; pd, pronephric duct; PLL, posterior lateral line; ter, terminal neuromasts. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)