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. 2016 Oct 13;113(1):15–29. doi: 10.1093/cvr/cvw218

Figure 3.

Figure 3

CD47 is required for hypoxia-mediated induction of pulmonary ET-1/ETA. (A) Age-matched male WT (CD47+/+), TSP1−/− and CD47−/− mice were challenged with normoxia (Nx) or short-term hypoxia (Hx) (10% FiO2, 24 h). Lung tissue lysates from WT and TSP1−/− mice (n = 3–5 per group) were prepared and protein separated via SDS–PAGE electrophoresis. Densitometry is presented as the mean ratio of target protein to α-tubulin ± SD; **P < 0.01, ***P < 0.001, ****P < 0.0001. Whole lung mRNA expression of TSP1 and CD47 from Nx or acutely Hx WT, TSP1−/−, and CD47−/− mice. Data from n = 3–5 mice per group are presented as mean ± SD, **P < 0.01. (B) Age-matched male WT and CD47−/− mice were challenged with Nx or chronic Hx (10% FiO2, 3 weeks). Lung tissue homogenates were prepared and protein separated by SDS–PAGE. Representative western immunoblots from treatment groups (n = 5–12 per group) for indicated proteins are presented. Densitometry is presented as the mean ratio of target protein to β-actin ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ET-1 levels in lung homogenates from WT and CD47−/− mice under Nx or chronic Hx were measured by ELISA. Data are presented as mean ± SD from n = 4–8 samples per group, ***P < 0.001, ****P < 0.0001. (D) mRNA expression of TSP1, CD47, cMyc, ET-1, ETA, and ETB in whole lung homogenates. Data from n = 6–8 mice per group are presented as mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (E) Platelet poor plasma TSP1 levels from WT and CD47−/− mice under Nx or chronic Hx. Data are presented as the mean ± SD from n = 5–9 mice per group, *P < 0.05, **P < 0.01. (F) Myography results of proximal PA vessels from age-matched male WT mice treated with TSP1 (2.2 nM) and the indicated concentrations of acetylcholine (Ach), sodium nitroprusside (SNP), or endothelin-1 (ET-1). Results are the mean ± SD of n = 3–4 vessels per treatment, #P = 0.05, *P < 0.05, ***P < 0.001.