Fig. 6.

Increased expression of NF-κB p65 in the lung in response to zinc deficiency and CLP was found. Mouse lungs were inflated and fixed with formaldehyde immediately following lavage and perfusion. A: immunostaining was performed to detect p65 protein (magnification, ×200). Data are representative of a minimum of 5 mice per treatment group. B: 10 random images of each lung specimen were captured and analyzed. The number of brown pixels staining positive for total p65 protein was counted and compared between each treatment group (n = 5 per group; *Zn⁻/CLP vs. Ctrl/CLP, P < 0.05; ‡Zn⁺/CLP vs. Zn⁻/CLP, P < 0.05). C: immunofluorescent staining for p65 was also conducted in conjunction with confocal analysis to identify the cellular location of p65 (green) within lung parenchymal tissue. Rhodamine-labeled Ricinus communis agglutinin (RCA), a lectin-based stain for type I alveolar epithelial cell-specific RCA (red), as well as the nuclear stain 4′,6′-diamidino-2-phenylindole (DAPI; blue) were used to clarify the location, identity, and extent of p65-positive cells. Pink arrows designate p65-positive type I alveolar epithelia, whereas white arrows designate alveolar macrophages. HPF, high-powered field.