Fig. 5.

Role of the TMEM16A in the effect of RvE1 on the ASL height and the cilia beating phenotype. (A) Representative orthogonal views of confocal microscopy z-stacks showing the ASL for a CF sample in control condition (vehicle) and treated with RvE1 (10 nM). ASL is labeled using dextran Texas red and cells using calcein green. (B) Quantification of ASL height for CF hNEC from three donors (F508del homozygous), in control condition, after RvE1(10 nM) treatment and after a combination of RvE1 and Ani 9 (10 µm), a specific inhibitor of the TMEM16A channel added 30 min before RvE1. The dashed line at 7 μm illustrates the ASL height measured in non-CF hNEC. (C) The role of TMEM16A on cilia beating was investigated by treating the hNEC from two CF donors (F508del homozygous), with Ani9 30 min before the addition of RvE1. Points and ellipses corresponding to CF vehicle (blue), RvE1 10 nM (green), and Ani9+RvE1 10 nM (purple) groups. Times correspond to treatment duration. PERMANOVA tests were performed for Ani9+RvE1 versus control (vehicle). Statistical tests were performed by including all principal components. (D) Corresponding values of CBF, synchronization, and orientation for the two groups over time. Mean ± SEM; *P < 0.05 and **P < 0.01.