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. 2017 Nov 2;4(4):229–239. doi: 10.1016/j.gendis.2017.10.004

Figure 2.

Fig. 2

Sox9 down-regulates BMP2-induced expression of Smad7 in MSCs. (A & B) Western blotting analysis of Smad7 expression at protein level. C3H10T1/2 cells were infected with the indicated adenoviral vectors. At 7 and 11 days after infection, total cell lysate was prepared and subjected to Western blotting analysis with an anti-Smad7 antibody or anti-β-actin control (A). The Western blotting results were quantitatively analyzed (B). BMP2-induced Smad7 expression was higher than that in the GFP group (p < 0.05). In the BMP2+Sox9 group, overexpression of Sox9 downregulated BMP2-induced Smad7 expression at days 7 and 11 compared to that of the BMP2 group (p < 0.05). (C) qPCR analysis of Smad7 expression at mRNA level. C3H10T1/2 cells were infected with the indicated adenoviral vectors. Total RNA was isolated at 3, 7, and 11 days after infection and subjected to quantitative RT-PCR (qPCR) using Smad7-rescific primers. BMP2 upregulated Smad7 mRNA levels at days 3, 7, and 11 compared to the GFP group (p < 0.05). At days 7 and 11 overexpression of Sox9 decreased BMP2-induced Smad7 mRNA level, compared to that of the BMP2 group (p < 0.05). Gapdh was used as a reference gene. “*” p < 0.05, vs. Ad-GFP; “#” p < 0.05, vs. Ad-BMP2.