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. 2023 Dec 14;14:7918. doi: 10.1038/s41467-023-43587-3

Fig. 4. Generation of xenogametes by iPGCT.

Fig. 4

a Schematic diagram for iPGCT using Gobiocypris rarus (Gr) as the donor and Tg(cmv:mCherry) zebrafish (Danio rerio, Dr) as the host. b At the 1-cell stage, Gr embryos were injected with zebrafish-derived 9GMs (50 pg per mRNA) to induce iPGCs. GFP-UTRnanos3 was used to label iPGCs. The embryos were photographed at 90% epibody. c Time-lapse of Gr_iPGCs migration to the genital ridges of Dr after transplantation. d Fluorescent image of conventional PGCT embryos and iPGCT embryos at 36 hpf. Gr_PGCT_Dr, Gr PGCs transplanted to PGC-depleted Dr embryos; Gr_iPGCT_Dr, 9GMs induced Gr iPGCs transplanted to PGC-depleted Dr embryos. e Success rate of iPGCT and conventional PGCT at 1 dpf. Each point represents an independent experiment (n ≥ 2), and at least 77 embryos were manipulated per experiment. A representative example of three replicate is shown. All data are presented as mean values ± SEM. Two-tailed Student’s t-test was used to calculate the P values. f Fluorescence imaging of GFP-UTRnanos3 positive Gr-derived iPGCs (Gr_iPGCs) in Tg(cmv:mCherry) zebrafish host. g Immunofluorescence detection of Vasa-positive germ cells in dnd_MO1 injected Dr (Dr dnd_MO1), Gr, and Gr_iPGCT_Dr testes at 60 dpf. h Morphology of the sperm of Gr, Dr and Gr_iPGCT_Dr. i Gr_iPGCT_Dr sperm generated by Dr hosts produced F1 and F2 generations of Gr normally.