rabbit anti mouse pro spc  (Abcam)

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, Staining, Derivative Assay

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: Durable murine honeycombing varies with expression of Muc5b in the bleomycin, but not H1N1, models. HC were morphometrically quantified via unbiased stereology; initially a 350 micron grid was used to quantify the volume fraction of the cysts. Cystic areas were then randomly sampled using a 100 micron grid to determine cyst size. Lung volume was calculated using the Cavalieri method. (A) No significant differences were observed in cyst volume or number based on genotype among mice infected with H1N1. (B) Bleomycin-injured mice expressing Muc5b under control of the SPC promoter had a significant increase in cyst volume (P < 0.05). The overall number of cysts is independent of genotype (*P < 0.05, **P < 0.01, and ***P < 0.005). KO = knockout; WT = wild-type (see text for additional strain details).

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: Expressing, Infection, Knock-Out