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. Author manuscript; available in PMC: 2019 Jun 18.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2014 Jul 24;34(9):2086–2094. doi: 10.1161/ATVBAHA.113.303017

Figure 3.

Figure 3.

CD44 and osteopontin (OPN) protect human sclerotic valve interstitial cells (VICs) from calcification induced by bone morphogenetic protein 4 (BMP4). A, Bar graph representing fold change in gene expression of OPN and alkaline phosphatase (ALP) in aortic valve sclerotic VICs after 12 days of in vitro treatment of BMP4 (100 ng/mL). B, Osteopontin quantification by real-time polymerase chain reaction (RT-PCR) of AVSc-derived VICs under BMP4 treatments at 0, 3, 6, and 12 days. C, Western blotting of protein extracts from AVSc-derived VICs treated with BMP4 (100ng/mL) for 3, 6, and 12 days. Antibodies against CD44, OPN, and GAPDH were used. D, Quantification of Western blotting. E, RT-PCR for CD44v6 and GAPDH of total RNA extracted from AVSc-derived VICs treated with BMP4 (100 ng/mL). F, RT quantification. G, Bar graph represents calcium deposition on aortic valve sclerotic VICs after 12 days of treatment with BMP4 (100 ng/mL), neutralizing antibody (NAb) OPN (5 μg/mL), NI-IgG (5 μg/mL), and their combinations. *P=0.05. H, Alizarin red staining representing calcium deposition (red) on VICs after 12 days treatment with BMP4 (100 ng/mL), NAb CD44 (5 μg/mL), NAb OPN (5 μg/mL), and their combination.