Figure 2
Figure 2. Siglec-F and sialylated Siglec-F ligands are up-regulated upon OVA challenge. (A) Serial sections of frozen lung from WT OVA-sensitized and -challenged mice were stained with antibodies against MBP (left panel, reddish brown color is positive) or Siglec-F (right panel, blue color is positive). Only the inflamed lungs were positive, as shown. (B) Recombinant soluble Siglec-F-Fc was used to probe for Siglec-F ligands in the lungs from OVA-sensitized and -challenged (OVA) or OVA-sensitized and PBS-challenged (No OVA) mice. Positive staining appears a dark reddish-brown color. The arginine-mutated R114A Siglec-F-Fc was used as a negative control, as it is deficient in sialylated ligand binding. Results shown are typical of n = 4 for each group and representative of 2 experiments. (C) Higher-magnification photomicrograph of an OVA-sensitized and -challenged lung section, probed with Siglec-F-Fc. Bronchiolar cells of the lung epithelia (white arrowheads) and mononuclear cells in the lung parenchyma (black arrowheads) were positive for Siglec-F ligands. For panels A-C, a 10×/0.25 DPlan dry objective lens was used to visualize images, and an Olympus BH2 camera was used to capture them. (D) Surface area of the Siglec-F ligand–positive bronchiolar epithelia. Mouse lungs were immunostained with Siglec-F-Fc and the area of bronchial epithelial Siglec-F-Fc immunostaining was quantitated by image analysis, with results expressed in squared micrometer/micrometer length of the basement membrane of the bronchus. WT mice challenged with OVA had a significant increase in levels of Siglec-F-Fc epithelial immunostaining compared with control non–OVA-challenged WT mice. (E) Mouse lungs were immunostained with Siglec-F-Fc, and the number of positive peribronchial cells quantitated was by image analysis. WT mice challenged with OVA had a significant increase in the numbers of peribronchial Siglec-F-Fc–positive cells compared with control non–OVA-challenged WT mice. (D-E) ***P < .001. Each error bar represents the standard error of the mean (SEM).

Siglec-F and sialylated Siglec-F ligands are up-regulated upon OVA challenge. (A) Serial sections of frozen lung from WT OVA-sensitized and -challenged mice were stained with antibodies against MBP (left panel, reddish brown color is positive) or Siglec-F (right panel, blue color is positive). Only the inflamed lungs were positive, as shown. (B) Recombinant soluble Siglec-F-Fc was used to probe for Siglec-F ligands in the lungs from OVA-sensitized and -challenged (OVA) or OVA-sensitized and PBS-challenged (No OVA) mice. Positive staining appears a dark reddish-brown color. The arginine-mutated R114A Siglec-F-Fc was used as a negative control, as it is deficient in sialylated ligand binding. Results shown are typical of n = 4 for each group and representative of 2 experiments. (C) Higher-magnification photomicrograph of an OVA-sensitized and -challenged lung section, probed with Siglec-F-Fc. Bronchiolar cells of the lung epithelia (white arrowheads) and mononuclear cells in the lung parenchyma (black arrowheads) were positive for Siglec-F ligands. For panels A-C, a 10×/0.25 DPlan dry objective lens was used to visualize images, and an Olympus BH2 camera was used to capture them. (D) Surface area of the Siglec-F ligand–positive bronchiolar epithelia. Mouse lungs were immunostained with Siglec-F-Fc and the area of bronchial epithelial Siglec-F-Fc immunostaining was quantitated by image analysis, with results expressed in squared micrometer/micrometer length of the basement membrane of the bronchus. WT mice challenged with OVA had a significant increase in levels of Siglec-F-Fc epithelial immunostaining compared with control non–OVA-challenged WT mice. (E) Mouse lungs were immunostained with Siglec-F-Fc, and the number of positive peribronchial cells quantitated was by image analysis. WT mice challenged with OVA had a significant increase in the numbers of peribronchial Siglec-F-Fc–positive cells compared with control non–OVA-challenged WT mice. (D-E) ***P < .001. Each error bar represents the standard error of the mean (SEM).

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