Journal: American Journal of Respiratory Cell and Molecular Biology
Article Title: Muc5b Enhances Murine Honeycomb-like Cyst Formation
doi: 10.1165/rcmb.2019-0138LE
Figure Lengend Snippet: Wild-type mice develop Muc5b-filled honeycomb cysts (HCs) in response to lung injury. Antibodies used for immunohistochemistry included goat anti-mouse Muc5b (11765, 1:1000; Everest), rabbit anti-mouse pro-SPC (ab90716, 1:100; Abcam), and chicken anti-mouse Krt5 (Poly9059, 1:1000; BioLegend). Appropriate fluorescent secondary antibodies were obtained from Jackson ImmunoResearch, reconstituted at 1 mg/ml, and used at 1:200. (A) Muc5b protein is increased in response to productive influenza infection (28 d postinfection) or repeated bleomycin treatment (*P < 0.05 and ***P < 0.005). (B) Influenza A H1N1 (H1N1) infection does not lead to significant fibrosis compared with UV-irradiated control virus as measured by hydroxyproline, whereas repeated bleomycin injury leads to significant increases in hydroxyproline compared with saline-treated control (***P < 0.005). (C) H1N1 generates considerable airspace remodeling resembling honeycombing (Krt5 stained DAB) 75 days after infection (top left). Similar to what is observed in humans, H1N1 HCs are characterized by Krt5-basal cells surrounding dense mucociliary structures filled with Muc5b+ colloid and generally exclude type II pneumocytes (top right). HCs also develop in mice subjected to repeated bleomycin injury (bottom left). Repeat bleomycin injury–derived HCs are generally indistinguishable from H1N1-derived HCs (bottom right). Honeycomb structures are highlighted by red (DAB panels) and white (IF panels) asterisks. (D) Cysts in mice subjected to single-dose bleomycin persist in the absence of significant fibrosis. (E) In general, resolution of H1N1 injury involves more robust honeycombing than repeat or single-dose bleomycin.
Article Snippet: Antibodies used for immunohistochemistry included goat anti-mouse Muc5b (11765, 1:1000; Everest), rabbit anti-mouse pro-SPC (ab90716, 1:100; Abcam), and chicken anti-mouse Krt5 (Poly9059, 1:1000; BioLegend).
Techniques: Immunohistochemistry, Infection, Irradiation, Control, Virus, Saline, Staining, Derivative Assay