Fig 2: Tfcp2l1 is strongly expressed in primate PGCs.A, heatmap showing the expression of Blimp1, Tfcp2l1, and Tfap2c during cynomolgus monkey PGC development. B, heatmap showing the expression pattern of the indicated genes during human PGC development. C, schematic diagram of human iPSCs transforming into PGCLCs. D, phenotype of human iPSCs and iMeLCs. The scale bar represents 100 µm. E, immunofluorescence staining of Blimp1 (green) and Tfap2c (red) in human PGCLCs. The nuclei were counterstained with Hoechst 33342 (Hoechst). The scale bar represents 100 µm. F, qRT-PCR analysis of the expression levels of PGC markers in human iPSCs transformed into PGCLCs in the presence or absence of PGCLC-inductive cytokines. Data are the mean ± SD (three independent experiments). *p < 0.05, **p < 0.01 versus without cytokines. Blimp1, B lymphocyte–induced maturation protein-1; cyESC, cynomolgus monkey ESCs; hPGC Wk9 F, week 9 female human PGCs; hPGC Wk9 M, week 9 male human PGCs; iPSCs, induced pluripotent stem cells; PGCd43, PGC on day 43; PGCLCs, PGC-like cells; PGCs, primordial germ cells; Tfap2c, transcription factor AP-2 gamma; Tfcp2l1, transcription factor CP2-like 1.

Fig 3: Tfap2c inhibits c-Myc-dependent apoptosis.a Microscopy of reprogramming cells at post-infection day 4, and the number of OSKM and OSKMT reprogramming cells at different time points was recorded. Scale bar, 100 µm. b Immunostaining of Ki67 in OSKM and OSKMT groups at day 4. Scale bar, 50 µm. c Flow-cytometer analysis of cell apoptosis after infected with OSK, OSKT, OSKM, and OSKMT on day 4. d The percent of Annexin V+ cells in OSK, OSKT, OSKM, and OSKMT group on day 4. e Immunoblotting of Cleaved caspase-3 (CC3) and Cleaved PARP1 (CPARP1) in MEFs after infected with OSK, OSKT, OSKM, and OSKMT on day 4. f Immunoblotting of CC3 and CPARP1 in MEFs after infected with OSK, OSKT, OSKM, and OSKMT on day 8. g Immunoblotting of CC3 and CPARP1 in MEFs after infected with OSKM and OSKMT (day 4) with or without PAC-1 (1.25 µM) treatment. h Number of Oct4-GFP+ colonies (day 12) in reprogramming cells treated with or without PAC-1. Significance in panels d was determined by one-way ANOVA with Dunnett’s test. Significance in panels a and h was determined by two-way AVOVA with Sidak’s multiple comparisons test. Significance summary: P > 0.05 (ns); *P = 0.05; ***P = 0.001. All data are presented as mean ± S.D., N = 3.

Fig 4: Primordial germ cell-like cells (PGCLCs) form at the interface between extraembryonic and primitive streak-like cells.(A, B) Immunofluorescence for different marker genes (maximal intensity projection along z). Yellow arrowhead in (B) points to higher NANOG expression in PGCLCs than pluripotent cells in the colony center. (C) Segmentation of nuclei based on DAPI staining. (D–F) Scatterplots of marker expression colored for radial position, normalized to threshold and log(1 + x) transformed, (D) corresponds to (A); (E, F) correspond to (B). (G) Scatterplot of PRDM1 vs. SOX17 colored for TFAP2C. (H, I) Spatial distribution of positive cells, dark lines represent the mean kernel density estimate of the positive fraction over four colonies, colored bands represent the standard deviation. (J) Clusters generated by Louvain. (K) Heatmap of differential expression between clusters (average z-scores) of genes associated with gastrulation. (I) PHATE visualization of scRNA-seq data showing denoised expression of markers used in (B–I) (raw in Figure 1—figure supplement 2D). (M) Scatterplot of TFAP2C vs. SOX17 from denoised scRNA-seq data (raw in Figure 1—figure supplement 1P), with colors matching clusters in (J). Scale bars 50 µm. All colonies are 700 µm diameter.

Fig 5: Short-term effects of TFAP2C, GADD45B, and PMAIP1 expression levels on NSCLC cell proliferation. a The effects of TFAP2C overexpression, GADD45B knockdown, or PMAIP1 knockdown on cell proliferation of WI-26 VA4 cells were measured by using cell viability assays. *p < 0.05 compared to cells treated with control vector or control siRNA. b–e The effects of TFAP2C, GADD45B, and/or PMAIP1 knockdown on cell proliferation of NCI-H292 (b), NCI-H358 (c), NCI-H460 (d), or A549 (e) cells were measured by using cell viability assays. *p < 0.05 compared to cells treated with control siRNA, **p < 0.05 compared to cells treated with TFAP2C siRNA