Journal: Nature Communications
Article Title: Non-lytic clearance of influenza B virus from infected cells preserves epithelial barrier function
Figure Lengend Snippet: IBV survivor cell populations are predominantly composed of cells expressing ciliated cell markers. a Tracheas were collected from Mock and Mal/04-Cre infected mice at 2 and 14 DPI then opened longitudinally and flattened to image the surface area of the whole-mount tracheal epithelium. Scale bar = 25 μm. b Quantification of apical surface area covered by tdTomato+ survivor cells from whole-mount trachea images. n = 4, 12 images per group, respectively, Student’s T -test. c Flow cytometry of 14 DPI tdTomato- (uninfected cells) and 14 DPI tdTomato+ (survivor cells) from the lungs of Mal/04-Cre infected mice, where cells are stained with CD24 (ciliated), SSEA-1 (secretory), GSIB4 (basal) cell surface markers. d Quantification of flow cytometry. n = 3 mice per group. e Cross-sectioned microscopy of murine tracheal epithelial cells from mice 14 days post-infection with Mal/04-Cre stained with KRT5 to label basal stem cells and SSEA-1 to label secretory cells. f Cross-sectioned microscopy of murine tracheal epithelial cells from mice 14 days post-infection with Mal/04-Cre stained with FOXJ1 to label ciliated cells. g Quantification of tdTomato+ survivor cells stained with cell-specific markers: KRT5, SSEA-1, and FOXJ1 compiled from two independent experiments. Data are presented as the number of marker+ cells out of total tdTomato+ cells and the percentage was calculated for marker+ cells out of total tdTomato+ cells imaged. S.E.M was plotted to indicate variability around the mean for tested samples, * p ≤ 0.05, ** p ≤ 0.001, ns not significant
Article Snippet: Samples were then stained with Actub (Abcam, clone EPR16772, cat ab179484, 1:1000), Krt5 (Biolegend, clone Poly19055, cat 905501, 1:500), FoxJ1 (eBioscience, clone 2A5, cat 14-9965, 1:500), SSEA-1 (Biolegend, clone MC-480, cat 125611, 1:250) ZO-1 (ThermoFisher, clone ZO-1-1A12, cat 33-9100, 1:500), CD24 (Abcam, clone M1/69, cat ab64064, 1:250) (Supplementary Table ).
Techniques: Expressing, Infection, Mouse Assay, Flow Cytometry, Cytometry, Staining, Microscopy, Marker