Fig 4. Inhibition of Na⁺ permeation through ENaC and NCX channels slowed PAEC scratch wound closure.

(A) Brightfield images of scratched PAEC monolayers at 0, 6, and 24-hours in vehicle control, 90 μM amiloride, and 10 μM benzamil conditions. Benzamil treatment caused accumulation of floating particles over 24 hours. Studies were all conducted in Na⁺-containing media. Scatter plots of wound closure distance in (B) 0–6, (C) 6–24, and (D) 0–24 hours. Benzamil was more potent than amiloride to slow PAEC monolayer wound closure. Data were reported as mean ± SEM. (E) Magnified view of the marginal regions of the control, amiloride-treated, and benzamil-treated cells after 24 hours of migration. An arrowhead points to a floating particle. Statistical significance was assessed using one-way ANOVA with Bonferroni post hoc test (ns—not significant, *—P < 0.05, **—P < 0.01, ***—P < 0.0001).