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. 2021 Jun 17;57(6):2002774. doi: 10.1183/13993003.02774-2020

FIGURE 2.

FIGURE 2

Nasal epithelial cultures derived from patients homozygous for other class II cystic fibrosis transmembrane conductance regulator (CFTR) mutations exhibit low CFTR function and variable levels of rescue by VX-445+VX-661+VX-770. DMSO: dimethyl sulfoxide; amil: amiloride; FSK: forskolin; CNX: calnexin; Ieq: transepithelial current; WT: wild-type. a–f) Representative tracings show Ussing chamber measurements of CFTR function in nasal epithelial cell cultures from CF patients bearing a) M1101K/M1101K, c) G85E/G85E and e) N1303K/N1303K in the absence or presence of the small-molecule corrector. Immunoblots of steady-state expression of WT and b) M1101K, d) G85E and f) N1303K following treatments with CFTR modulators. Markers in kDa. Band C: mature, complex-glycosylated CFTR; band B: immature, core-glycosylated CFTR. g) Maximal response Ieq after stimulation by FSK (10 µM)±VX-770 (1 µM) for nasal epithelial cell cultures from one to three technical replicates of three patients bearing M1101K or G85E and two donors bearing N1303K after pre-treatment (48 h at 37°C) with DMSO (0.1%), VX-809 (3 µM)+VX-770 (1 µM), R-VX-445 (3 µM)+VX-661 (3 µM)+VX-770 (1 µM) or S-VX-445 (3 µM)+VX-661 (3 µM)+VX-770 (1 µM). h) Ieq by the CFTR inhibitor CFTRinh-172 (10 µM) for nasal epithelial cell cultures from one to three technical replicates of three patients bearing M1101K or G85E and two donors bearing N1303K. i) Comparison of IeqCFTRinh-172 across the class II mutations M1101K, G85E and N1303K for nasal epithelial cell cultures from one to three technical replicates of three patients bearing M1101K or G85E and two donors bearing N1303K. Dashed line in g–i) represents the mean results of rescued (r) F508del/F508del nasal cells treated with VX-445+VX-661+VX-770. j) Ratio band C/(band C+band B) from one technical replicate of three patients bearing M1101K or G85E and two patients bearing N1303K. Data are presented as mean±sd. Analysis was performed using one-way ANOVA followed by Turkey's post hoc test (*: p<0.05; **: p<0.01; ***: p<0.001; ****: p<0.000). The paired t-test showed statistically differences between nasal epithelial cells treated with DMSO and VX-809+VX-770 (#: p<0.05; ##: p<0.01). ns: nonsignificant.