Figure 4.

Inhibition of ANO1 Cl⁻ channels by Ani9. (A) ANO1‐YFP fusion protein is targeted to the plasma membrane of HEK293 cells. The cell surface is labeled with the lectin WGA, conjugated to a fluorescent dye. (B) Representative traces of Ca²⁺‐induced Cl⁻ currents in Ano1‐YFP‐transfected HEK293 cells with and without the Cl⁻ ‐channel blockers T16A _inh ‐A01 and Ani9. Whole‐cell currents were recorded at ‐70 mV with 0.75 μmol/L Ca²⁺ in the pipette solution following formation of the whole‐cell configuration (arrow). (C) Statistical summary of experiments in HEK293 cells, illustrating that the blocking efficiency depended on the intracellular Ca²⁺ concentration. Data are means ± SEM (n = 9–17 HEK293 cells per value). (D) Whole‐cell current‐to‐voltage relation of ANO1‐mediated Cl⁻ currents at 0.75 μmol/L intracellular Ca²⁺ and block by 10 μmol/L extracellular Ani9. Currents from 10 to 12 HEK293 cells were averaged, and background currents at 0 Ca²⁺ were subtracted.