Fig. 6 | Nature Communications

Fig. 6

From: Non-lytic clearance of influenza B virus from infected cells preserves epithelial barrier function

Fig. 6

Survivor cells contribute to the maintenance of epithelial barrier integrity during IBV infection. a CD24+ (ciliated) cells were isolated from Mock and Mal/04-Cre infected murine lungs at 14 DPI and forward scatter (as a measure of cell volume) was measured for mock infected cells, tdTomato− (uninfected cells) and tdTomato+ (survivor cells). b Quantification of the geometric mean of cell forward scatter, n = 3 mice per group, Student’s T-test. c Microscopy of flattened whole-mount tracheal epithelium at 9 DPI with survivor cells expressing tdTomato (red) stained with ZO-1 (green) for tight junctions. Scale bar = 10 μm. d Quantification of whole-mount microscopy cell size according to ZO-1 staining for tdTomato− (uninfected cells) and tdTomato+ (survivor cells) in the same image. n = 10 images, with at least four cells quantified per group in each image, Student’s T-test. e Quantification of whole-mount microscopy cell size according to ZO-1 staining for ciliated tracheal cells from mock infected mice and tdTomato+ (survivor cells) from Mal/04-Cre infected mice. n = 21 uninfected ciliated cells, eight survivor cells from three images each. Student’s T-test. fi WT and DTR-transgenic mice were either mock or Mal/04-Cre infected and treated with diphtheria toxin (DT) to ablate survivor cells specifically in the DTR-transgenic mice). f Epithelial permeability was measured by administering 4 kDa fluorescein isothiocyanate (FITC)-Dextran and measuring leakage into the surrounding tissue at 11 DPI, 2 days after depletion of survivor cells. n = 4, 3, 3 mice per group, respectively, one-way ANOVA. g Albumin leakage into the bronchoalveolar lavage fluid (BALF) was measured by ELISA at 11 DPI, 2 days after depletion of survivor cells. n = 5 mice per group, one-way ANOVA. h Static compliance of lungs was measured using Flexivent physiology studies at 11 DPI, 2 days after depletion of survivor cells. n = 5, 5, 7 mice per group, respectively, two-way ANOVA. i Body weight curves of tdTomato and DTR-transgenic mice infected and treated with DT. n = 5, 5, 4 mice per group, respectively, one-way ANOVA at each timepoint. S.E.M. was plotted to indicate variability around the mean for tested samples, *p ≤ 0.05, **p ≤ 0.001, ns not significant

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