Fig 2: Combinatorial and individual roles of TFAP2C, SOX17 and PRDM1 in epigenetic regulation of target genes in hPGCLCs.a, The intersection of TFAP2C, SOX17 and PRDM1 peaks in hPGCLC aggregates. Statistical significance of overlap was determined by hypergeometric test.b, The enrichment of TFAP2C, SOX17 and PRDM1 peaks in promoters and enhancers that became active or inactive during the PreME to hPGCLC transition (see Extended Data Fig. 5d). The TF peaks were categorized into seven cooperativity classes as in a. Dot size represents the fraction of enhancers/promoters that overlapped with the TF peaks.c, The direct up target genes of TFAP2C, SOX17 and PRDM1. The heatmaps show the expression of representative target genes during hPGC development (left) and the expression pattern in TFAP2C (day 2), SOX17 (day 2) and PRDM1 (day 4) knockout (KO) hPGCLCs/aggregates versus wild-type control (CTL) (middle). The representative gene ontology terms enriched in the direct target genes based on the binding cooperativity of TFAP2C, SOX17 and PRDM1 are shown on the right.d, Genome browser snapshots of representative TFAP2C, SOX17 and PRDM1 direct up target genes. e, The direct down target genes of TFAP2C, SOX17 and PRDM1 and the representative gene ontology terms.f, Enrichment of NANOG, POU5F1, TEAD4 binding sites (ReMap2020 non-redundant peaks) in TFAP2C, SOX17 and PRDM1 peaks in hPGCLCs.g, The enhancer-linked TF network that establishes the hPGC program.

Fig 3: Blimp-1 negatively regulates cell migration in SCC but not HaCaT cells. After transfection of silencing Blimp-1 and control shRNA, HaCaT (A), SAS (B) and Cal-27 (C) cells were seeded in wound-healing assay kit and grown overnight for attachment. Then the kits were removed and fresh DMEM medium with mitomycin C (5 µg/ml) was treated. After 30 min, cells were stimulated with PMA (30 nM), TNF-a (10 ng/ml) or EGF (50 ng/ml). Photography (100x) was taken by microscopy. Dashed lines represent boundaries of the wounds. The percentage of wound closure from the denuded gap after incubation for different times was determined. *p < 0.05 (mean ± S.E.M., n = 3), indicating the significant enhancement effects of shBlimp-1 and EGF, and the inhibitory effect of PMA on cell migration as compared to control cells.

Fig 4: IL-21 upregulates Blimp-1 in an ERK-dependent manner. BxPC-3 and Panc-1 were incubated with IL-21 (10 and 25 ng/mL) for 24 h. (A) Treatment with IL-21 upregulates Blimp-1 expression. One of six independent experiments is shown. (B) Quantification of IL-21-dependent Blimp-1 upregulation (IL-21: 10 ng/mL). * P < 0.05. Wilcoxon rank-sum test. (C) ERK inhibitor prevents IL-21-dependent Blimp-1 upregulation. One of five independent experiments is shown.

Fig 5: OHL cells express both KLF4 and BLIMP1.H&E analysis (Image: 40X), and immunohistochemistry analysis was performed on a paraffin-embedded, formalin-fixed biopsy of an oral hairy leukoplakia (OHL) lesion using antibodies directed against Z, BMRF1, KLF4 and BLIMP1 as indicated (Images: 100x of region boxed in H & E stain). Examples of OHL cells, positively staining for each of these proteins, are indicated by black arrows.