Figure 2.
Figure 2. Regulation of Nogo-B expression during asthma-like conditions in mice and human asthma. (A) Nogo-B protein levels were significantly decreased in total lung lysates of OVA-sensitized and challenged (allergic) mice versus WT control (nonallergic) mice. Shown is a typical Western blot from six individual experiments. Nogo-B levels were quantified by densitometry and normalized to hsp90 (*, P < 0.05). Data are expressed as mean ± SEM. n = 4–7 mice per group. (B) Lung sections from representative small and large airways of untreated (control, a and b) and OVA-challenged (c and d) WT mice stained with Nogo-A/B antibody. Brown indicates positive Nogo-A/B in airway epithelium (arrowheads), smooth muscle and vessels (asterisk) of control mice, and the loss of Nogo immunoreactivity in epithelium and smooth muscle, but not vessels (*, c and d), of OVA-challenged mice. Bar, 100 µm. The graph on the right depicts quantification of Nogo-A/B staining with multiple sections from multiple mice using the scoring system, where 3 is 100% positive staining and 0 is an absence of staining (*, P < 0.01). Data are expressed as the mean ± SEM. n = 4–8 mice for both groups and data are representative of three experiments. Serial sections were stained with E-cadherin (e and f), H&E (g and h), and PAS (i and j). (C) Representative images of large (a and b) and small (c and d) airways from human lung sections of normal and fatal asthmatic specimens stained for Nogo with the N-18 antibody. Airway epithelium is indicated with arrowheads and vessels with asterisks. Bars: (top) 200 µm; (bottom) 100 µm. Images are representative of four individual normal human lung specimens and one individual fatal asthmatic specimen, with additional patient samples in Fig. S2. (D) Lung sections from control (top) and OVA-treated (bottom) Nogo-A/B+/− LacZ mice stained for β-gal. Airways (both E-cadherin and SMA positive) are indicated with A and vessels (SMA positive only) are indicated with V. Bars, 100 µm. Images represent results obtained from three individual experiments where n = 6 mice.

Regulation of Nogo-B expression during asthma-like conditions in mice and human asthma. (A) Nogo-B protein levels were significantly decreased in total lung lysates of OVA-sensitized and challenged (allergic) mice versus WT control (nonallergic) mice. Shown is a typical Western blot from six individual experiments. Nogo-B levels were quantified by densitometry and normalized to hsp90 (*, P < 0.05). Data are expressed as mean ± SEM. n = 4–7 mice per group. (B) Lung sections from representative small and large airways of untreated (control, a and b) and OVA-challenged (c and d) WT mice stained with Nogo-A/B antibody. Brown indicates positive Nogo-A/B in airway epithelium (arrowheads), smooth muscle and vessels (asterisk) of control mice, and the loss of Nogo immunoreactivity in epithelium and smooth muscle, but not vessels (*, c and d), of OVA-challenged mice. Bar, 100 µm. The graph on the right depicts quantification of Nogo-A/B staining with multiple sections from multiple mice using the scoring system, where 3 is 100% positive staining and 0 is an absence of staining (*, P < 0.01). Data are expressed as the mean ± SEM. n = 4–8 mice for both groups and data are representative of three experiments. Serial sections were stained with E-cadherin (e and f), H&E (g and h), and PAS (i and j). (C) Representative images of large (a and b) and small (c and d) airways from human lung sections of normal and fatal asthmatic specimens stained for Nogo with the N-18 antibody. Airway epithelium is indicated with arrowheads and vessels with asterisks. Bars: (top) 200 µm; (bottom) 100 µm. Images are representative of four individual normal human lung specimens and one individual fatal asthmatic specimen, with additional patient samples in Fig. S2. (D) Lung sections from control (top) and OVA-treated (bottom) Nogo-A/B+/− LacZ mice stained for β-gal. Airways (both E-cadherin and SMA positive) are indicated with A and vessels (SMA positive only) are indicated with V. Bars, 100 µm. Images represent results obtained from three individual experiments where n = 6 mice.

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