The results indicated that the mRNA expression of SMAD2 and SMAD5 in inhibition groups was significantly suppressed compared with the control group during chick germ cell differentiation in vitro and in vivo ; furthermore, their protein expression was consistent with mRNA expression before and after inhibition, while DNA methylation showed a decreasing trend during germ cell differentiation, implying that the TGF-β/BMP4 signaling pathway could promote male germ cell formation and that DNA methylation may regulate this process (see Figure 3 ).
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DNA Methylation and Regulatory Elements during Chicken Germline Stem Cell Differentiation.
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