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. 2019 Dec 20;7:351. doi: 10.3389/fcell.2019.00351

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Copyright © 2019 Gao, Zhang, Dai, Zhang, Li and Bai.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Specific genes were significantly up-regulated during stage III of beta cell differentiation from MSCs. (A) Schematic diagram of the differentiation program. The program was divided into five stages according to the order in which the induction factors were added. (B) Flow cytometry analysis of the expression of Ngn3 and Pdx1 during each stage of differentiation. (C) Immunofluorescence microscopy was used to determine the expression of Pdx1 and insulin during each stage of differentiation. (D) Western blotting was used to analyze the expression of proteins during each stage, including the islet hormones, Ngn3 and Pdx1, and the main members of the TGF-β/Smad pathway, namely p-Smad2, p-Smad3, and Smad4. The data showed that these proteins were markedly elevated from stage III, implying that the TGF-β/Smad pathway plays an important role in beta cell differentiation. Values represent the mean ± SEM, n = 3. The paired two-tailed t-test was used for comparisons (^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001). Gapdh and Histone were used as endogenous controls in the lysate and nuclear fractions, respectively.