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Fig. 7. Sorting and functional assay of L812AL813A-ClC-2-HA mutant Cl^– channel. (A) MDCK cells were grown to confluence upon class coverslips, transfected with ClC-2-HA (MDCK::ClC-2) or L812AL813A-ClC-2-HA. Cells were observed 24 hours post-transfection after staining with an anti-HA antibody. Specimens were analyzed by confocal microscopy. Representative X-Y and X-Z sections are shown. Calibration bar: 10 µm, applies to all panels. (B) Representative current traces in a HEK-293 cell transfected with L812AL813A-ClC-2. V_H was 0 mV test pulses ranged from –170 to 0 mV as indicated. These were followed by a pulse to 30 mV. The duration of the main pulses was increased at more positive voltages in order to approximate full activation of the conductance. For illustration proposes, the beginning of the tail currents at 30 mV were set at the same time. (C) Apparent conductance calculated by taking the current at the beginning of the pulses at 30 mV given after various conditioning prepulses to the voltages in the abscissa. Results are means±s.e.m. of four separate experiments. Triangles, data from L812AL813A-ClC-2-HA-transfected cells. Circles, data for WT ClC-2, taken from Niemeyer et al. (Niemeyer et al., 2004).

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