(A) Heatmap showing normalized expression of indicated genes in PGCs, AGCs and hESCs, H9 primed, H9 naïve and ICM. M=male, F=Female (B,C,F,G,I). Note that DNMT3L is enriched in the H9 naïve cells relative to the rest of the data sets where as SOX2 and UHRF1 are enriched in H9 primed cells. Representative immunofluorescence micrographs of UHRF1 (B,C,I), and DNMT3A (F,G,I) with germline markers cKIT or VASA in prenatal testes (B,F) and ovaries (C,G) at the developmental stage indicated in days and with pluripotency marker OCT4 in UCLA1 hESCs (I). Arrows indicate UHRF1 or DNMT3A signal. (D,E) Quantification of UHRF1 in cKIT+ or VASA+ germ cells in testes (D) and ovaries (E), at the developmental ages indicated days (d). (D) In testes for quantification in cKIT+, 14 optic fields were counted at the PGC stage from n=4 testes. For the AGC stage, 23 optic fields were counted from n=3 testes at 87–95 days and 28 optic fields from n=4 testes at 105–119 days of development. For quantification in VASA+, 12 optic fields were counted at the PGC stage from n=4 testes. For the AGC stage, 22 optic fields were counted from n=3 testes at 87–95 days and 25 optic fields from n=4 testes were counted at 105–119 days of development. e, In ovaries for quantification in cKIT+, 14 optic fields were counted at the PGC stage from n=3 ovaries. For the AGC stage, 13 optic fields were counted from n=3 ovaries at 70–95 days, 9 optic fields from n=3 ovaries at 105–116 days and 8 optic fields from n=2 ovaries at 126–130 days of development. For quantification in VASA+, 13 optic fields were counted at the PGC stage from n=3 ovaries. For the AGC stage, 15 optic fields were counted from n=3 ovaries at 70–95 days, 8 optic fields from n=3 ovaries at 105–116 days and 8 optic fields from n=2 ovaries at 126–130 days of development. For immunofluorescence microscopy, nuclei were counterstained with DAPI (blue). Scale bars, 10 um. Data are represented as mean±sem. Days (d), neg (negative). (H) Expression levels of TET1-3 in prenatal testes and ovaries from 53 days to 137 days (n= number of samples at each developmental stage). Also see Figure S5.