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. 2013 May 10;8(5):e63138. doi: 10.1371/journal.pone.0063138

Figure 4. BMP-9-induced SMAD1/5/8 phosphorylation is impaired in both 2f/1f- and 1f/1f-ECs.

Figure 4

Western blot showing SMAD1/5/8 phosphorylation, total SMAD1 protein, and β-actin in response to various concentrations of BMP-9 (A, B) and BMP-4 (C, D). A. Comparison of SMAD1/5/8 phosphorylation induced by BMP-9 shows no significant difference between 2f/1f and 1f/1f derived from 2f/1f-ECs. B. 2f/2f-ECs are 10-fold more sensitive to BMP-9 than 1f/1f-ECs derived from 2f/2f, suggesting that BMP-9-SMAD1/5/8 signaling is impaired in the 1f/1f-ECs. C and D. There is no difference in SMAD1/5/8 phosphorylation in response to BMP-4 in both 2f/1f (C) and 1f/1f (D) ECs. β-Actin was used as a loading control. Histogram shows phospho-SMAD1/5/8 normalized by total SMAD1 and β-Actin relative to the value indicated by asterisks. Data in all graphs represent means of values measured by densitometry from three separate blots. Error bars are SDs (*p<0.05 and **p<0.01).