Fig. 2: Chloride and GSH permeabilities of the LRRC8/VRAC conductance measured in WT and LRRC8A-KO HEK293 cells.

a Whole-cell Cl⁻ currents recorded in WT (left) and LRRC8A-KO HEK 293 cells (right), under control conditions (Iso, 340 mOsm l⁻¹), in the presence of DCPIB (10 µM, Iso + DCPIB) and after replacing the bath with a hypotonic solution (Hypo, 270 mOsm l⁻¹). Once the Cl⁻ conductance was fully developed (3–4 min), DCPIB was perfused. b Whole cell currents recorded as in (a) after replacement of the NMDGCl pipette solution with a GSH pipette solution. Bath solution compositions are the same as in (a) containing NMDG⁺/Cl⁻ as major ions. c Mean current/voltage relationships measured in HEK293 WT recorded after the stabilization of the VRAC Cl⁻ current (hypo). Current values were measured 6–10 ms after the onset pulse. The means I/V relationship recorded under asymmetrical pipette and bath solution is also plotted (GSH pipette solution versus Cl bath solution). Note that all the relationships illustrated are corrected by the junction potential calculated for each experimental condition. d, e Box plots illustrating the remaining GSH current (d, measured at −120 mV) and the GSH conductance (e, calculated between −100 and −60 mV) and their sensitivity to DCPIB exposure (20 µM) at −120 mV. Currents were recorded with GSH pipette solution and hypo NMDGCl bath solution as in (b). Records were obtained from 13 to 14 individual cells (Mann–Whitney and Wilcoxon paired tests were used, ***p < 0.001).