Fig. 5
From: Phenotypic profiling of CFTR modulators in patient-derived respiratory epithelia
ACC assay reports primary defect and pharmacological rescue of major CF mutant in lung cultures differentiated from iPSCs. a ES cell from Wt (CA1) and iPSC derived from F508del CF patient (GM00997) were differentiated to airway epithelia as previously described.47 ACC is measured in these epithelia. Airway tissue generated from Wt CA1 ES cells show a robust response to CFTR stimulation by FSK using the fluorescence-based detection method and the analysis is shown in Fig. 1. The changes in fluorescence in activated cultures were normalized to fluorescence measurements in vehicle (DMSO) treated cultures. The traces (upper panel) are representative of three biological replicates (or three transwells) wherein >50 regions within each transwell were monitored over time. iPSC-derived airway epithelium from CF-affected individuals were rescued with corrector VX-661 (1 µM) or DMSO as control (lower panel). CFTR channels were activated in all cultures by FSK and VX-770 (1 μM). b Bar graphs represent maximum percentage change in fluorescence (ΔF) after addition of CFTR agonist, relative to baseline (F0) measurements prior to agonist addition (time = 10 min). Asterisks indicate statistical significance using one-way ANOVA and Tukey’s multiple comparison tests for the three distinct differentiations (*p < 0.05, **p < 0.01, ***p < 0.001). Immunofluorescence images of CFTR expression in non-CF ES cells and CF iPS cells are shown in Supplementary Fig. 10
