Fig. 2: Quantification of FIS in iPSC-derived airway epithelial spheroids from non-CF donors.
From: A multimodal iPSC platform for cystic fibrosis drug testing
A Schematic of FIS assay with representative images of two spheres immediately prior to as well as 6, 12, 18, and 24 h after forskolin addition. B Kinetics of non-CF airway spheroid FIS measured by the change in cross-sectional area (CSA). Top panel shows individual spheroid FIS (black lines) and average FIS (green lines) (n = 3 experiments). The middle panel shows compiled FIS (and vehicle control) experimental averages with magnified inset indicating 20–24 h. Bottom panel shows mean and standard error for each time point depicted in the middle panel (n = 3 experiments). C Experimental approach for the FIS assay. Each iPSC line was differentiated in three independent experiments and FIS performed on day 28–32 (n = 3 biological replicates of independent differentiations were analyzed per donor for each of three donors). Representative images of non-CF airway epithelial spheroids (day 28) are shown on the right. D Automated imaging analysis of non-CF spheroids before (left) and after (right) forskolin stimulation using OrganoSegTM to quantify the change in CSA. E Number of airway spheroids analyzed per experiment for the non-CF cell line indicated. Each point represents an independent experiment. F Three non-CF donor FIS responses shown. Each point represents an independent experiment. No significant differences between samples, by one-way ANOVA. Scale bars represent 250 µm. Lines and error bars represent mean ± standard error.
