Fig. 2: N3 suppresses the induction of hPGCLCs from hESCs.
From: RNA-binding proteins DND1 and NANOS3 cooperatively suppress the entry of germ cell lineage
A Venn diagram showing the overlap of D1-interacting proteins28 and hPGCLC-specific genes40. B Reciprocal co-immunoprecipitation showing the physical interaction of D1 and N3 in hESCs. Representative images from three independent experiments. Source data are provided as a Source Data file. C The morphology of control and N3-KO hESCs. Scale bar: 200 μm. D RT-qPCR showing relative expression levels of N3 in control hESCs, control and N3-KO day 4 hPGCLC aggregates. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. E Flow cytometry showing the percentages of day 4 hPGCLCs induced from control and N3-KO hESCs. F Statistics of hPGCLC percentages from control and N3-KO hESCs. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. G Immunofluorescence of OCT4 (cyan), SOX17 (magenta) and PRDM1 (yellow) in control and N3-KO day 4 hPGCLC cryosection. Scale bar: 20 μm. Representative images from three independent experiments. H RT-qPCR showing relative expression levels of SOX17, PRDM1 and TFAP2C in control hESCs, control and N3-KO day 4 hPGCLC aggregates. Mean with SD from n = 3 experiments. Unpaired t test; two-tailed P value. Source data are provided as a Source Data file. I Flow cytometry showing the percentages of hPGCLCs induced from control and N3-KO + N3-GFP hESCs. J Statistics of hPGCLC percentages from control, N3-KO (GFP negative) and N3-KO + N3-GFP (GFP positive) 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. K PCA plot showing the transcriptomes of control, D1-KO, N3-KO hESCs, and sorted control, D1-KO, N3-KO day 4 hPGCLCs, gonadal hPGCs8, and hESC-derived endoderm, mesoderm, and ectoderm. L Plot showing the DEGs of control and N3-KO day 4 hPGCLCs. M Heatmap showing the expression of pluripotency and PGC marker genes in control, D1-KO, N3-KO hESCs, and sorted control, D1-KO, N3-KO day 4 hPGCLCs. See also Supplementary Fig. 2.
