1. Deleterious impact of hyperglycemia on cystic fibrosis airway ion transport and epithelial repair 
Claudia Bilodeau, Olivier Bardou, Émilie Maillé, Yves Berthiaume, Emmanuelle Brochiero 
Journal of Cystic Fibrosis 
Volume 15, Issue 1, Pages (January 2016)
DOI: /j.jcf
Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

2. Fig. 1
Impact of high glucose exposure on Cl− currents through non-CF and CF CFBE airway cell monolayers. CFBE-wt and CFBE-ΔF508 cell monolayers were cultured on permeant filters at the air-liquid interface and treated for a 24-h period on the basolateral side with low (5mM glucose+20mM mannitol, LG) or high (5+20mM glucose, HG) glucose, prior to short-circuit current (Isc) measurements in an Ussing chamber. Apical and basolateral sides were then bathed with a normal physiological solution. Basal total Isc current and transepithelial resistance (TER) were then measured in LG and HG conditions (B, n=8), before application of 10μM amiloride (to inhibit ENaC currents) and then, forskolin (10μM) and IBMX (100μM) to stimulate cAMP-activated CFTR channels, followed by the CFTRInh172 (20μM, apical, see a representative trace in A). Quantification of the mean ΔIFk+IBMX and ΔICFTRInh172 short-circuit currents (in μA/cm2) after LG and HG exposure are reported in panels C and D (n=8). E and F. UTP (100μM, apical) was applied to stimulate calcium-activated Cl− currents (see a representative trace in E), while mean ΔIUTP currents are reported in F (n=7). ⁎: P<0.01.
Journal of Cystic Fibrosis  , 43-51DOI: ( /j.jcf )
Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

3. Fig. 2
Reduction of basolateral K+ currents through CFBE airway cell monolayers by hyperglycemia. CFBE-wt and CFBE-ΔF508 cell monolayers were cultured on permeant filters at the air-liquid interface and treated for a 24-h (B) or 48-h (C) period on the basolateral side with low or high glucose, prior to short-circuit current measurements in an Ussing chamber. In these experiments, filters were bathed with asymmetrical physiological media (high K+ at the apical side and low K+ at the basolateral side, see Materials and methods). The apical membrane was then permeabilized with 7.5μM amphotericin B (Ampho B). Upon stabilization of the current, clofilium (100μM) and then glibenclamide (100μM) were applied at the basolateral side (see representative Isc trace in A). Total basolateral K+ currents after apical membrane permeabilization with amphotericin B (ΔIAmphoB, B, C) as well as clofilium- and glibenclamide-sensitive K+ currents (ΔIclofi+glib, B) were then compared under LG and HG conditions, through CFBE-wt (n=14) and CFBE-ΔF508 (n=6–7). ⁎: P<0.04.
Journal of Cystic Fibrosis  , 43-51DOI: ( /j.jcf )
Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

4. Fig. 3
Decreased wound-healing rates by hyperglycemia in non-CF and CF CFBE airway cell monolayers. CFBE-wt and CFBE-ΔF508 cell monolayers were exposed to a normal culture medium (5mM glucose, Ctl, B), a culture medium supplemented with mannitol (20mM mannitol+5mM glucose, LG, B, C, D, E) or glucose (25mM glucose, HG, B, C, D, E) for 24h (C, n=12), 48h (D, n=17) or 7days (B, n=8; E, n=12) before wounding. The wound-closure rates (μm2/h) were monitored over a 6-h period (see representative photographs presented in A) and then compared in control and LG conditions (B) and in LG and HG conditions (C, D, E). ⁎: P<0.02.
Journal of Cystic Fibrosis  , 43-51DOI: ( /j.jcf )
Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

5. Fig. 4
Decreased wound-healing rates by hyperglycemia in rescued CF airway cell monolayers. CFBE-ΔF508 (A, n=13; B, n=7) and primary human CF airway (C, n=8) epithelial cell monolayers were exposed to culture medium containing low or high glucose, in the absence or presence of the CFTR corrector VRT-325 (5μM) for 24h before wounding and/or using chamber measurements. The wound-closure rates (μm2/h) were then monitored over a 6-h period. ⁎: P<0.03. For Isc measurements, a Cl− gradient was established by bathing the cell monolayers with an asymmetrical physiological solution (low Cl− concentration solution at the apical side and high Cl− concentration at the basolateral side) and the basolateral membrane was permeabilized with 7.5μM amphotericin B. Cells were then sequentially exposed to a combination of forskolin (10μM, Fk), IBMX (100μM) and genistein (30μM, Gen), followed by CFTRInh−172 (20μM). The mean ΔIFk+IBMX+Gen and ΔICFTRInh172 short-circuit currents (in μA/cm2) were measured in LG, LG+VRT-325 and HG+VRT-325 conditions (B).
Journal of Cystic Fibrosis  , 43-51DOI: ( /j.jcf )
Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions