Fig. 5 | Nature Communications

Fig. 5

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

Fig. 5

Ciliated-like survivor cells display reduced expression of ciliated cell transcription factors and fail to elaborate surface cilia. a Quantitative RT-PCR measuring RNA levels of Forkhead Box J1 (Foxj1) in flow sorted EpCam+ CD24+ mock infected cells, tdTomato− (uninfected cells), and tdTomato+ (survivor cells) at 14 days post-infection. n = 3 mice per sample, one-way ANOVA. b Air–liquid interface differentiated cultures of primary murine epithelial cells were infected with Mal/04-mNeon (green) and stained for FOXJ1 (red). White arrows = mNeon+ (infected cells), yellow arrows = mNeon− (uninfected cells). c Quantification of FOXJ1 brightness per cell, normalized to brightness of uninfected cells. n = 6 images, at least four cells quantified per image for each group, Student’s T-test. d Microscopy of cross-sectioned murine tracheas from mock infected mice where epithelial cells were stained for FOXJ1 and acetylated tubulin for mature cilia. White arrow = tdTomato− (uninfected cells) with intact cilia. e Close-up panel of uninfected murine tracheal ciliated cells stained for FOXJ1 and ACTUB. White arrow = tdTomato− (uninfected cells) with intact cilia. f Quantification of uninfected cells that are double positive for FOXJ1 and ACTUB. g Microscopy of cross-sectioned murine tracheas from Mal/04-Cre infected mice at 14 DPI where epithelial cells were stained for FOXJ1 and ACTUB. White arrow = tdTomato− (uninfected cells) with intact cilia, yellow arrows = tdTomato+ (survivor cells) lacking surface cilia. h Close-up panel of murine tracheal uninfected Mal/04-Cre infected cells stained for FOXJ1 and ACTUB. White arrow = tdTomato− (uninfected cells) with intact cilia, yellow arrows = tdTomato+ (survivor cells) lacking surface cilia. i Quantification of Mal/04-Cre infected cells: tdTomato− (uninfected cells) and tdTomato+ (survivor cells) that are double-positive for FOXJ1 and ACTUB. 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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