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. 2018 Sep;59(3):383–396. doi: 10.1165/rcmb.2017-0127OC

Figure 3.

Figure 3.

Mucin-related phenotypes in lower airways of Spdef-deficient neonatal and adult mice. (A) Representative micrographs illustrating AB-PAS staining in neonatal (PND 10) naive Spdef-sufficient (Spdef+/+) and Spdef-deficient (Spdef−/−) mice. Insets show high-magnification views of areas highlighted by the arrows in the low-magnification panels. Micrographs are representative of at least three mice for each genotype. Scale bar: 200 μm. (B) Densitometric analysis of Muc5b- and Muc5ac-specific signaling in agarose Western blots of BAL from naive Spdef+/+ and Spdef−/− mice at PND 10 (n = 5–6/genotype). (C) Representative micrographs illustrating AB-PAS (upper panel), Muc5b (middle panel), and Muc5ac (lower panel) staining in the proximal main stem bronchus (left lung) of naive Spdef+/+ and Spdef−/− mice at PND 42. Insets show high-magnification views of areas highlighted by the arrows in the low-magnification panels. Micrographs are montages acquired with a slide scanner as described in the Methods section of the main text. Scale bars: 100 μm. (D–F) Morphometric quantification of (D) AB-PAS+, (E) Muc5b+, and (F) Muc5ac+ volume densities in proximal, intermediate, and distal sections of the main stem bronchus in naive Spdef+/+ (open dots) and Spdef−/− (solid dots) mice at PND 42. Data are presented as mean ± SEM and were analyzed with an unpaired Student’s t test. (G) Densitometric analysis of Muc5b- and Muc5ac-specific signaling in agarose Western blots of BAL from naive Spdef+/+ (open dots) and Spdef−/− (solid dots) mice at 4–5 months of age (n = 3–4/genotype). Note that Muc5ac was undetectable for both genotypes in adult mice. (H) Quantitative RT-PCR results for Muc5b and Muc5ac transcripts in naive Spdef+/+ (open dots) and Spdef−/− (solid dots) mice at PND 42 (n = 7–9/genotype). (I) Tracheal mucociliary clearance measurements in naive Spdef+/+ (open bar) and Spdef−/− (solid bar) adult mice (n = 9/genotype) at ages PND 43 (n = 7) and PND 170 (n = 2) for both genotypes. (J) Bioelectrical properties of tracheal epithelium isolated from naive Spdef+/+ (open bar) and Spdef−/− (solid bar) mice at 4–5 months of age (n = 5–7/genotype). *P < 0.05, analyzed with 2 tailed, unpaired Student’s t test.