Extended Data Fig. 1: Generation of ruminant haploid androgenetic embryos.

a. Schematic overview of haSCs derivation, haploid embryos were generated by removing female pronucleus from normal diploid zygotes. ♂, male pronucleus; ♀, female pronucleus. b. Immunostaining for 5-hydroxymethylcytosine (5hmC) in bovine (left) and ovine (right) embryos. The haploid androgenetic embryos are generated by sperm injection into enucleated oocytes or by removing female pronucleus from fertilized oocytes. The diploid IVF embryos were used as controls. ♂, male pronucleus; ♀, female pronucleus; n = 3 independent experiments with similar results. Scale bar, 25 μm. c. Immunostaining for trophectoderm marker CDX2 in the bovine and ovine blastocysts on day 7 after activation or insemination. The haploid and diploid embryos are generated as described in Extended Data Fig. 1b. n = 3 independent experiments with similar results. Scale bar, 50 μm. d. Quantification for the cell numbers in the bovine and ovine blastocysts on day 7 after activation or insemination. The haploid and diploid embryos are generated as described in Extended Data Fig. 1b; ICM, inner cell mass; Data are mean ± s.d. (n = 18 IVF, n = 19 injecting, n = 21 removing (top left); n = 18 IVF, n = 19 injecting, n = 21 removing (top right); n = 22 IVF, n = 19 injecting, n = 23 removing (bottom left); n represents total embryos of three independent experiments); P values are from unpaired, two-tailed Student’s t-tests.