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Caption : supplementary figure1 bmp4 and ra induced model differences affect the differentiation of esc. a. qrt-pcr results showed that the stra8 gene was induced by ra, but not by bmp4; b, c. simultaneous addition of bmp4 and ra in the medium was not conducive to the formation of embryoid bodies, scale bar: 50μm; d. qrt-pcr results showed bmp4 and ra have obvious antagonistic effects in the process of inducing germ cell differentiation.

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Caption : supplementary figure 2 dynamic changes of key molecules in bmp4 and ra signals during germ cell formation. a. dynamic expression of bmp4 in germ cell formation; b, c. expression of downstream signaling molecules and related target genes in bmp4 signaling pathway during germ cell formation; d. western blot assay results: bmp4 signal in pgcs activated during formation and inhibited during sscs formation. e. genes involved in ra synthesis: dynamic expression of adh5 and aldh1a1 during germ cell formation.

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Caption : supplementary figure 3 acetylation and dna methylation covalent modification of dazl involved in pgc differentiation. a. the pluripotency gene was inhibited during pgc formation in vivo; b. p300 gene was significantly up-regulated during pgc formation in vivo; c. in vitro acetylation assay showed that the level of intracellular acetylation was significantly up-regulated during pgc formation; qrt-pcr results showed that dazl gene was activated in bmp4 induction model; e. correlation analysis showed that dazl expression was regulated by bmp4 signal; f, g. dual luciferase assay showed that dazl promoter was protected by dna a co-modification of basalization and histone acetylation.

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Caption : supplementary figure 4 morphological changes of cells, stra8 promotes the formation of ssc induced by ra.

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Caption : supplementary figure 6 protein interaction analysis between bmp4 and ra signals. the results show that the interaction between ra and bmp4 signals can be through bmp4, aldh1a1, aldh1a2 and stub1 complexes.