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Figure S3: Validation of BMP pathway manipulation in HAECs. (A): Expression levels of pSMAD1/5 in response to BMP2 (100ng/ml, 2h), Noggin (100ng/ml, 2h), or BMP2+Noggin (100ng/ml each, 2h) in proliferating HAECs. Actin was used as a loading control. (B) Proliferating HAECs were stained to identify the specific sub-cellular localization of pSMAD1/5 (in green) in untreated control cells (panels 1-3), BMP2-treated cells (100ng/ml, 2h) (panels 4-6), Noggin-treated cells (100ng/ml, 2h) (panels 7-9) and cells treated with both BMP2 and Noggin (100ng/ml each, 2h) (panels 10-12). Nuclei were stained with DAPI (in blue; panels 1,4,7,10). Data are representative of 3 independent experiments. (C): Dose-response curve of Noggin treatment. Regenerating HAECs were chronically treated with different doses of recombinant Noggin. Acetylated-tubulin positive MCCs were counted at LC. 100ng/ml Noggin was the minimal dose to give the maximal effect and used in the rest of the study. Image published in: Cibois M et al. (2015) Copyright © 2015. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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