Fig. 8: Contribution of Japanese ptarmigan-derived iPSCs to chicken embryos.

a iPSCs were injected into chicken blastoderm after fluorescently labeling them with a cell tracer. b Interspecific blastoderm was generated after the injection of Japanese ptarmigan-derived iPSCs into chicken blastoderm. Images shows embryos that were not injected (i–iv) and iPSC-injected blastoderm (v–viii). Panels show the merged bright field and green fluorescence images (i, v), bright field images (ii, v), green fluorescence (iii, vii), and red fluorescence (iv, viii). Bars represent 2 mm. c The images show the contribution of iPSCs to the chicken embryo post-injection after 72 h. Arrows indicate green signals. Panels show embryos with no injection (i–iv), and iPSC-injected blastoderm (v–viii). Panels show merged bright field and green fluorescence images (i, v), bright fields (ii, vi), green fluorescence (iii, vii), and red fluorescence (iv, viii). Bars represent 2 mm. d Reaction of anti-hygromycin antibody with Japanese ptarmigan iPSCs. Staining images of the anti-hygromycin antibody with red fluorescence-labeled secondary antibody (i–iii), staining images of the anti-hygromycin antibody with green fluorescence-labeled secondary antibody (iv–vi). Panels show merged images (i, iv), green fluorescence (ii, v), and red fluorescence (iii, vi). Scale bar represents 50 μm. e Detection of GFP, hygromycin resistance gene, and DAPI signals in embryonic cryosections on day 5. Panels show the merged image (i, v), the GFP (ii, vi), the hygromycin resistance gene (iii, vii), and the DAPI (iv, viii) stained images. Bars represent 50 μm (upper) and 100 μm (lower). f Detection of reprogramming vector cassette from Japanese ptarmigan-injected chicken embryos on day 5. The electrophoretic image of the detection of the reprogramming vector (PB-TAD-7F) and the detection of the internal control (Tsc-2) shows. g, h Detection of reprogramming vector cassette from Japanese ptarmigan-injected chicken embryos on day 5 with real-time PCR. g Amplification plots of PB-TAD-7F (reprogramming vector) and Tsc-2 in chicken embryo (injection minus), Japanese ptarmigan iPSCs, and chimera No. 9 (iPSC-injected chicken embryo No. 9). h Quantification of PB-TAD-7F in iPSC-injected chicken embryos. Centerlines of box plots indicate medians; box limits indicate the 25th and 75th percentiles. n = 6.