Fig. 2: AAV6 diffusion in control and IL-13 treated mucus from in vitro cultures.

A Schematic illustration showing the different in vitro models from which mucus was collected. BCi-NS1.1 cells were grown at ALI to establish Healthy ALI cultures. Healthy differentiated ALI cultures were stimulated with IL-13 (10 ng/ml) for 7 days to produce MUC5AC rich mucus. MUC5B KO cultures were generated using lentiviral mediated delivery of sgRNA and CRISPR-Cas9. Mucus from healthy controls, IL-13 treated cultures and MUC5B KO cultures were collected and AAV6 diffusion was measured in comparison to muco-inert nanoparticles of similar size. B Scatter plots of measured median log₁₀[MSD_1s] for AAV6 and 20 nm muco-inert nanoparticles in healthy mucus collected from ALI cultures. ****p < 0.0001 by Mann–Whitney Test. C Representative trajectories of AAV6 diffusion in control mucus, IL-13 treated mucus, and MUC5B KO mucus. Trajectory color changes with time with dark blue indicating 0 s and dark red indicating 5 s. Scale bar = 1 µm. D Scatter plots of measured median log₁₀[MSD_1s] for AAV6 in healthy control mucus, IL-13 treated mucus, and MUC5B KO mucus. p > 0.05 by Kruskal–Wallis test with Dunn’s correction. Each dot represents data from 1 video (n ≥ 3).