Fig. 3: D1 and N3 cooperatively restrict the induction of hPGCLCs from hESCs.

A RT-qPCR showing relative expression levels of D1 and N3 in control hESCs, control and D1-N3-dKO day 4 hPGCLCs. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. B FACS analysis showing the percentages of control and D1-N3-dKO day 4 hPGCLCs. C Statistics of hPGCLC percentages from control and D1-N3-dKO hESCs. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. D PCA plot showing the transcriptomes of control, D1-KO, N3-KO, D1-N3-dKO hESCs, and sorted control, D1-KO, N3-KO, D1-N3-dKO day 4 hPGCLCs, gonadal hPGCs⁸, and hESC-derived endoderm, mesoderm, and ectoderm. E Plot showing the DEGs of control and D1-N3-dKO day 4 hPGCLCs. F Heatmap showing the expression of pluripotency and PGC marker genes in control, D1-KO, N3-KO hESCs, and sorted control, D1-KO, N3-KO, D1-N3-dKO day 4 hPGCLCs. G FACS analysis showing the percentages of control (RFP negative) and D1-OE (RFP positive) day 4 hPGCLCs. Scale bar: 80 μm. H Statistics of hPGCLC percentages from control and D1-OE hESCs. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. I FACS analysis showing the percentages of control (GFP negative) and N3-OE (GFP positive) day 4 hPGCLCs. Scale bar: 80 μm. J Statistics of hPGCLC percentages from control and N3-OE hESCs. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. K FACS analysis showing the percentages of control and D1-N3-dOE day 4 hPGCLCs. Scale bar: 80 μm. L Statistics of hPGCLC percentages from control and D1-N3-dOE hESCs. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. See also Supplementary Fig. 3.