Figure 6.

Phosphorylation of Thr⁵³² in ClC‐3 was involved in AngII‐induced migration in A10 cells. A. The effect of ClC‐3 knockdown or ROCK inhibition on AngII (1 μmol·L⁻¹)‐induced wound healing in BASMC from wild type (WT) and ClC‐3 null (KO) mice. a. Representative images of wound healing assay carried out in control, AngII‐treated and Y27632‐ treated groups. AngII increased the wound healing (distance cells migrated), which was inhibited by Y27632 (10 μmol·L⁻¹) or ClC‐3 channel knockdown. b. Bar charts of relative wound healing from the experiments. n = 6, *P < 0.05 vs. WT con. B. The effect of ClC‐3 channel knockdown or ROCK inhibition on AngII(1 μmol·L⁻¹)‐induced transwell migration of BASMC from WT or ClC‐3 KO mice. a. Representative images of transwell assay carried out in control, AngII‐treated and Y27632‐ treated groups. AngII significantly increased the transwell rate, which was inhibited by Y27632 or ClC‐3 knockdown. b. Bar charts of relative migration rate from the experiments. n = 6, *P < 0.05 vs. WT con. C. The effect of Thr⁵³² phosphorylation on AngII(1 μmol·L⁻¹)‐induced wound healing in A10 cells. Representative images of wound healing assay in cells transfected with plasmids of N1,WT ClC‐3, T532A and T532D mutants. Cells were pretreated with or without Y27632 in N1‐transfected, WT ClC‐3‐transfected and T532D‐transfected groups. AngII‐induced wound healing was increased in cells transfected with WT ClC‐3 plasmid, which was inhibited in cells transfected with the T532A mutant and further potentiated in cells transfected with the T532D mutant. Y27632 markedly reduced wound healing in WT ClC‐3‐transfected cells, but had no effect on T532D‐transfected cells. D. The effect of Thr⁵³² phosphorylation on AngII(1 μmol L⁻¹)‐induced transwell migration in A10 cells. Representative images of transwell assays in cells transfected with plasmids of N1, WT ClC‐3, T532A and T532D mutants. Cells were pretreated with or without Y27632 in N1, ClC‐3 and T532D transfected groups. AngII‐induced transwell migration was increased in cells transfected with ClC‐3 plasmid, which was inhibited in cells transfected with T532A, and further potentiated in cells transfected with the T532D mutant. Y27632 markedly reduced the transwell migration in WT ClC‐3 transfected cells, but had no effect on T532D transfected cells. E. Bar charts of relative wound healing and relative migration from the experiments . n = 5, *P < 0.05 vs. con, #P < 0.05 vs. Ang.