Fig. 3

Elevating endogenic TGF-β/Nodal signal to promote DE differentiation. A Quantitative RT-PCR analyzed signaling pathway-related genes expression patterns in DE induction with different protocols. Data are presented as mean ± SEM, n = 3. B Immunostaining analyzed SOX17 and BRA expression in the CILy and CIP-derived differentiated cells. Scale bars 100 μm. C Representative immunoblots of NODAL, TGF-β1, and the phosphorylation of Smad2/3 at day 2 during DE induction with different protocols. D Quantification of the protein expression levels. Data were expressed as mean ± SD, n = 3 for each group, * P < 0.05, ** P < 0.01 vs control (CI) group