Fig. 4

PGCLC differentiation from pDFPs. A Scheme showing PGCLC and SSCLC induction from SDSCs. B Representative morphology of PGCLCs generated from pDFPs at increasing culture times. Pictures were taken using an eclipse microscope (10x, Nikon TE2000, Japan). Scale bars, 100 μm. C Expression of the germ cell marker DDX4 and DAZL in day 6 eGFP positive pDFP-derived PGCLCs. Pictures were taken using an confocal microscope (20x, Leica, TCS SP5 II, Germany). Scale bars, 25 μm. D Expression of genes proper of specified PGCs by day 2, 4, 6 pDFP-derived PGCLCs. E Western blotting analysis of H3K27me3 and H3K9me2 expression during PGCLCs differentiation from pDFPs. F Bisulfite sequencing of DMRs of the imprinted genes Igf2r and Peg3 in P2 SDSCs, day 6 pDFP-derived PGCLCs and E12.5 PGCs