Figure 2.

Lung progenitor cultures express WT CFTR and channel activity in fluorescence-based assay

(A) Representative FLIPR trace (mean ± SEM) responses of four wells plated with submerged cultures stimulated by 10 μM forskolin and inhibited by 10 μM CFTRInh-172. This cell line was derived from donor CF2MC for which the F508del mutation was corrected to WT. The method for mutation editing is described in Eckford et al. (2019), and characterization of the CF2MC line is given in Document S1. The naming of lines is consistent with Figure 1.

(B) Western blot shows molecular weight (MW) markers and mature CFTR expression in submerged culture of the same line, (180 kDa). Calnexin (CNX) was used as loading control.

(C) Top, heatmap shows representative data for a single plate (48 wells), used for establishing assay statistics (p value, Z-prime factor, and SSMD). Multiple wells were seeded with iPSCs differentiated to submerged lung progenitor cells. Alternating rows (each containing six wells) show well scans of CFTR channel activation after agonist (forskolin, FSK) or vehicle control (DMSO) addition. The response size is color coded as shown in the side bar, with red representing the highest response and purple-black, the lowest response. Bottom, bars show data from all of the above wells treated with control (DMSO) or FSK. Assay statistics are superimposed.