Extended Data Figure 5 |. AmB restores ASL pH and antibacterial activity in primary cultures of human airway epithelia from donors with CF.

(a) Genotypes and Δ pH measurements of patient donors in ASL pH assay. (b) The effects of AmB (2 μM, 48 hours; n = 9 biologically independent samples, p = 0.446), C35deOAmB (2 μM, 48 hours; n = 5 biologically independent samples, p = 0.9994) and basolateral addition of AmB (2 μM, 48 hours; n = 3 biologically independent samples, p = 0.6359) on the average ASL pH of primary cultured airway epithelia derived from CF humans with different CFTR mutations. (c) The effect of AmB (2 μM, 48 hours; n = 7 biologically independent samples, p = 0.4866) on ASL pH in non-CF epithelia. (d) Genotypes of patient donors in ASL viscosity assay. (e) Genotypes of patient donors in ASL antibacterial activity assay. (f) The effect of AmB (2 μM, 48 hours; n = 8 biologically independent samples, p = 0.0042) and C35deOAmB (2 μM, 48 hours; n = 5 biologically independent samples, p = 0.9626) on the average ASL antibacterial activity of primary cultured airway epithelia derived from CF humans with different CFTR mutations. Antibacterial activity is measured by the % of S. aureus killed after exposure to ASL. (g) The ability of AmB (2 μM) alone to kill S. aureus as compared to saline (n = 36 biologically independent samples, p = 0.1569). Representative FRAP traces for measuring ASL viscosity from 6 independent experiments repeated with similar results are shown in (h) non-CF, (i) CF, and (j) AmB-treated CF epithelia. In panels (b) and (f), ANOVA was used to assess statistical significance. In panel (c), two-sided unpaired Student’s t test with Welch’s correction was used. In panel (g), two-sided unpaired Student’s t test was used to assess statistical significance. Graphs depict means ± SEM; NS, not significant; *P ≤ 0.05; **P ≤ 0.01. In all panels, measurements were taken from biologically independent samples.