查看文章

Methods

To investigate DDR resulting from cigarette smoke in vivo, male C57BL/6 mice were challenged for 28 days with cigarette smoke or ambient air, as previously described (9). BOEC were isolated from human blood samples, and human primary bronchial epithelial cells were isolated from lung tissue as described (3, 10). Informed consent was obtained from all individuals (Table 1). Data have been derived from subjects and animals used in previous reports (3, 9, 10). Human umbilical vein endothelial cells were transfected with miRCURY locked-nucleic-acid microRNA inhibitor hsa-miR-126-3p (Exiqon;# 4109148-001) or negative control (# 199006-001) at 50 nM. RNA was extracted using the mirVana Isolation Kit (Ambion) or the RNeasy kit (Qiagen). miR-126-3p levels, and the small nucleolar RNAs RNU44 and snoRNA135 as reference genes, were measured by TaqMan assays (# 477887,# 4427975 …