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. 2019 Sep 17;9:13446. doi: 10.1038/s41598-019-49758-x

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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BMP9 peptide P3 is a potentiator for BMP9 signalling in PAECs. (A) SPR sensorgrams of peptide P1*, P2* and P3* binding to ALK1-Fc on CM5 chip. Raw data were fitted to steady state binding kinetics to obtain the K_D values of 25M, 84.7 M and 2 mM for P1*, P2* and P3*, respectively. (B) SPR sensorgrams of ALK1-Fc binding to peptide P3* on SA chip. The steady state fit yielded a K_D value of 30 mM. (C) Representative immunoblots against pSmad1/5, and total Smad1, of the protein extracts from PAECs that have been treated with BMP9 (at 0.03 ng/ml, or 1.24 pM growth factor domain (GFD) dimer) in the presence or absence of P3. Right: quantification of the pSmad1/5 bands by densitometry, N = 3. Paired t-test. (D) Effect of peptide P3 (at 160 μM) on BMP9 (at 0.3 ng/ml, or 12.4 pM GFD dimer)-induced gene expression of ID1, HEY1 and HEY2 following 1.5-hour treatment, or expression of BMPR2 following 5-hour treatment, in hPAECs. N = 6, data shown as means ± S.E., paired t-test.