Fig. 3: AtCLCf is involved in efflux of Cl⁻.

a Chloride-selective ion transport by MQAE fluorescence quenching assay showed transport of chloride ion by AtCLCf. Data are mean ± SD (n = 3). (Inset to a) Schematic representation of chloride-selective, MQAE dye-based fluorescence assay for ion transport. Liposomes without incorporation of AtCLCf were used as control. Arrow indicates the time point where different salts were added. The schematic was created using PowerPoint software. b AtCLCf showed increased transport of chloride with addition of increasing NaCl concentrations in MQAE based quenching assay. Data are mean ± SD (n = 3). c–e Enhanced inward and outward current was observed in cells expressing AtCLCf as compared to the vector transfected cells. c Exemplar traces of a vector transfected cell showing the current evoked from holding potential of 0 mV to different test potential ranging from −80 mV to 80 mV with 10 mV increment. d Exemplar trace of AtCLCf cell. e Current voltage relationship of respective groups. f, g Data comparing the maximal outward and inward current at 80 and −80 mV respectively. **P < 0.01 (e–g data are mean ± SE, vector n = 12, AtClCf n = 14). Statistics was performed with unpaired Student’s t test (2-tailed). h–j The effect of replacing most of internal CsCl with CsF, CsBr, CsI, CsNO₃ or CsAcetate on the ionic current. h Current voltage relationship of AtCLCf transfected cell recorded with 140 mM CsCl-based internal as compared to cell recorded with 130 mM CsF, CsBr, CsI, CsNO₃ or CsAcetate and 10 mM CsCl-based internal solution. The profiles for vector and AtCLCf cell recorded with CsCl based internal solution were reproduced from e for comparison, i and j data comparing the maximal outward and inward current at 80 and −80 mV respectively. (h–j data are mean ± SE, n = 12 for CsF, 11 for CsBr, 9 for CsI, 12 for CsNO₃ and 9 for CsAcetate). Different letters within a data set are significantly different, P < 0.01 (one-way ANOVA followed by Tukey’s test).