Fig. 1

Generation of transgenic zebrafish of Tg(nanos3:nfsB-mCherry-nanos3 3′UTR). (A) Schematic diagram showing the construct used to generate the transgenic zebrafish line of Tg(nanos3:nfsB-mCherry-nanos3 3′UTR). (B) Schematic diagram showing the work flow of setting up the transgenic zebrafish. (C) Genotyping results of F1 zebrafish by PCR amplification. The gel picture was cropped from an original full-length gel image provided in Supplementary information. The identities of PCR products were further confirmed by Sanger sequencing. (D–F) Photomicrographs of the typical F2 embryos at 24 hpf displaying the expression of mCherry at the genital ridge (D: light; E: fluorescence; F: merge). Embryos were observed laterally. (G) PCGs with fluorescence were arranged as two lines. Embryos were observed dorsally. (H) Standard curve comparison. Standard curves for transgene Tg(nano3:nsfb-mCherry nanos3 3′UTR) and the endogenous β-actin gene (actb) in serially diluted (10-fold) DNA samples from the cloning vector of Tg(nano3:nsfb-mCherry nanos3 3′UTR) and wild type zebrafish genome. A very efficient amplification was obtained, as indicated by the slopes of the standards curves. Ct, cycle threshold.