Figure 2.

Characterization of opened and 3D iPSC-HIOs. (a) Schematic depicting the generation of opened organoids by removal of the extracellular supporting matrix. (b) Gene expression RT-qPCR studies of CFTR and ENaC and SLC6A14 in opened organoids relative to 3D organoid expression. (c) Western blot of WT-CFTR expression in 3D and opened non-CF organoids, compared to expression in a Human Bronchial Epithelial (HBE) cell line. HBE cells expressing either WT-CFTR and or genetic knockout of CFTR as positive and negative controls for CFTR protein expression, respectively, as previously described [22]. (d) Immunofluorescence of CFTR (green), apical membrane marker, ZO-1 (red), and nuclei (blue) in opened non-CF01 organoids.