Fig. 8

Ccl2, S100a8, S100a9, and Saa3 expression is increased in Adgrf5^−/− lung ECs. a Cytokine and chemokine expression profile in lavage lungs from three mice (12-weeks-old), analyzed using a Proteome Profiler Mouse Cytokine Arrays as described in the Methods. The data from Adgrf5^−/− mice are expressed as values relative to those from WT mice. Values are presented as the average of duplicate measurements. Asterisks indicate CCL2 and IL-13. b–e The mRNA expression of Ccl2 (b), S100a8 (c), S100a9 (d), and Saa3 (e) was analyzed by qPCR in whole (18.5 days post-coitum and 2 days and 1 week of age) or post-lavage (3, 6, and 30 weeks of age) lungs from WT and Adgrf5^−/− mice. The data were normalized to Gapdh levels and are expressed as values relative to those from corresponding WT mice. Values are presented as the mean ± SEM (n = 3–6). *p < 0.05; **p < 0.01; ***p < 0.001. f The mRNA expression of S100a8, S100a9, Saa3, and Ccl2 was analyzed by qPCR in primary lung ECs from WT and Adgrf5^−/− mice (1-week-old). The data were normalized to Gapdh levels and are expressed as values relative to those from WT mice. Values are presented as the mean ± SEM (n = 4–5). *p < 0.05 and **p < 0.005. g Whole cell lysates (10 μg of protein) from lung ECs were analyzed by western blotting using antibodies specific for CCL2 (top), ADGRF5 (middle), and α-tubulin (bottom). The band intensity of CCL2 was normalized to that of α-tubulin and expressed as a value relative to that from WT mice. Values are presented as mean ± SEM (n = 5). *p < 0.05