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heximide-sensitive fashion. Identification of a specific mRNA for (via Northern blotting) and the presence of immunoreactive species in cultured RFLETII cells after cytokine, LPS, and ZAS exposure provide unambiguous evidence of the localization of within this key pulmonary cell type. The extent of *NO production by epithelium, which exceeds that of other lung cell types, reveals new insight into mechanisms of pulmonary host defense and pathways of free radicalmediated lung injury. EXPERIMENTAL METHODS Materials. Minimum essential medium (MEM), Hanksâ balanced salt solution (HBSS), and antibiotic/antimycotic solution were from GIBCO Laboratories (Grand Island, NY). Heat-inactivated fetal calf serum (FCS) was from Hyclone Laboratories (Logan, UT). Recombinant murine interleukin- 1 p (IL- 1 p>, interferon-y (IFN-$, and tumor necrosis factor-a (TNF-CX) were from R & D Systems (Minneapolis, MN). Cycloheximide, Escherichia coZi 011 l:B4 LPS, L-NMMA, and zymosan were from Sigma Chemical (St. Louis, MO). Timed-pregnant female Sprague-Dawley rats were from Charles River Laboratories (Wilmington, MA). CeZZ culture. RFLE-TII cells were isolated and cultured as previously described (6). Briefly, 19- or 20-day-gestation fetuses were aseptically removed from their mothers, and the lungs were dissected out, minced, and resuspended in cold HBSS (calcium and phosphate free). The minced tissue was trypsinized
AJP - Lung Cellular and Molecular Physiology – The American Physiological Society
Published: Mar 1, 1995
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