Figure 6.

Expression of CFTR protein in W57G/A234D by western blot analysis

Rectal OGs (upper panel), nasal epithelial cells (lower panel). Non-CF controls are made of intestinal organoids (HC) or Calu3 cells, respectively.

(A) Densitometric analysis of the results shown as representative western blots in (B). The controls show the presence of C and B bands related to the different CFTR glycosylation states. F508del/F508del samples were used as a reference and show an increase in the expression of B and C bands following treatment with a combination of VX661 with VX445. The newly synthesized non-glycosylated polypeptidic chain (band B) is expressed at variable levels in W57G/A234D colonoids. Treatment for 24 h of intestinal colonoids with CFTR modulator VX809, VX661 alone, or in combination with VX445 improved the expression of band C reaching statistical significance. In VX809-treated rectal OGs, the increased expression almost reaches statistical significance (p = 0.06). An overlapping trend of drug-induced CFTR Band C increase was observed in nasal cells. β-actin polyclonal antibody was used as a loading control. The results of the densitometric analysis are shown in (A), summarize the β-actin-normalized data (C + B bands) from a minimum of three independent experiments. ∗p < 0.05, ∗∗p < 0.005 and ∗∗∗p < 0.002 (ANOVA and Kruskal–Wallis tests).