Figure 5
From: Somatic cell hemoglobin modulates nitrogen oxide metabolism in the human airway epithelium
Hemoglobin β is colocalized with endothelial nitric oxide synthase (eNOS) in normal human airway epithelial cells. (A) Immunofluorescent staining of Hbβ (green) and eNOS (red) are co-localized (yellow) in primary airway cells NHE grown on ALI, permeabilized, fixed and labeled as described in the methods. Studies were performed by confocal microscopy, Leica, 60X. (B) Immunohistochemical staining of Hbβ in an endobronchial biopsy from a healthy subject in the SARP. Normal lung biopsy revealed presence of Hbβ at the apex of airway epithelial cells (arrow) and some in inflammatory cells, × 40 magnification. (C) Colocalization of eNOS and Hbβ in normal human airway epithelial cell cultures. Cell cultures at ALIs were lysed in RIPA buffer and immunoprecipitated (IP) for Hbβ followed by immunoblot (IB) for and eNOS (left panel); in separate cells, proteins were IP’d for eNOS followed by IB for Hbβ (right panel). Immunoprecipation and IB were performed as described in the methods, total. N = 5.
