Supplementary Material
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Supplementary material. Methods and tables.ERJ-00419-2019.SUPPLEMENT
Supplementary figure S1. Deposition of Tc99-DTPA in ferret tissues. a) Representative uncorrected images of Tc99-DTPA deposition in the lungs acquired at baseline (Time 0). b) Total deposition of radiolabel at Time 0 was quantified before assessment of clearance. c) Systemic absorption of Tc99-DTPA as monitored by uptake into forearm muscles.ERJ-00419-2019.Fig_S1
Supplementary figure S2. Schematic of experimental protocol for ferret trachea. Upon euthanisation, the trachea was removed from each animal and immediately dissected for μOCT imaging, followed by cholinergic stimulation of mucus secretion for assessment of mucus properties such as microrheology and percent solids by weight. Thick borders indicate endpoint collection throughout the protocol. Capitalised words indicate key timepoints for endpoints. Abbreviations: μOCT: micro-optical coherence tomography, PTM: particle tracking microrheology.ERJ-00419-2019.Fig_S2
Supplementary figure S3. Distribution of μOCT-quantified airway epithelial function by region of interest (ROI). μOCT was used to measure a) ASL depth, b) PCL height, c) CBF, and d) MCT rate at multiple ROI per ferret trachea, to account for the local microenvironment. Data is presented as box-and-whisker (Tukey method). n=1078 videos.ERJ-00419-2019.Fig_S3
Supplementary figure S4. Cigarette smoke exposure reduces MCT in primary HBE cells. a) Representative μOCT stills of primary HBE cells after 24-h apical treatment with 2% cigarette smoke extract (CSE) or DMSO control. Red line: ASL. Yellow line: PCL. Quantification of in vitro ASL (b) and MCT (c) following exposure. n=10 ROI; **: p<0.01, ***: p<0.001 compared to vehicle control, as determined by unpaired Mann–Whitney (ASL) and t-test (MCT).ERJ-00419-2019.Fig_S4
Supplementary video S1. Representative μOCT video of mucus transport in a trachea excised from an air control ferret.ERJ-00419-2019.Video_S1
Supplementary video S2. Representative μOCT video of mucus transport in trachea from a smoke-exposed ferret.ERJ-00419-2019.Video_S2