Axon pclamp 10.7 software suite

Single currents were recorded using cell-attached configuration, as described previously [63 (link)]. Briefly, the patch-clamp set-up was based on the Axopatch 200B operational patch-clamp amplifier (Molecular Devices, San Jose, CA, USA) and Axon Digidata 1550A (Molecular Devices, San Jose, CA, USA) analog–digital converter controlled by a Windows-based personal computer with installed Axon PClamp 10.7 Software Suite (Molecular Devices LLC, San Jose, CA, USA) for data acquisition, processing and analysis. Patch pipettes were pulled from borosilicate glass capillaries (BF-150-86-10, Sutter Instruments, Novato, CA, USA) to a resistance of 10–15 MOhm when filled with the extracellular solution containing (in mM): 145 NaCl, 2 CaCl₂, 1 MgCl₂ and 10 HEPES/TrisOH. Potassium bath solution containing 145 KCl, 2 CaCl₂, 1 MgCl₂ and 10 HEPES/TrisOH was used to nullify the resting membrane potential of the cells. The pH of all solutions was maintained at 7.3. All experiments were performed at RT. The single-channel recordings were processed and analyzed in Clampfit software (part of Axon PClamp 10.7 Software Suite). Single-channel conductance values were defined by the slope of the current–voltage relationship (I–V) after linear approximation. Averaged conductance values are given as the mean ± SEM (n—number of experiments).

The patch–clamp setup was based on the Axopatch 200B operational amplifier and Axon Digidata 1550A (Molecular Devices LLC, San Jose, CA, USA) analog-to-digital converter, controlled by a Windows-based personal computer with Axon PClamp 10.7 Software Suite (Molecular Devices LLC, San Jose, CA, USA) for data acquisition, filtration, processing and analysis. Pipettes were pulled from borosilicate glass capillaries with filament (BF-150-86-10, Sutter Instruments, Novato, CA, USA) to a resistance of ~5–10 MΩ when filled with a cytosol-like solution (see Section 2.3). All experiments were performed at room temperature (22–23 °C). Whole-cell currents were recorded using a whole-cell configuration of the patch–clamp technique, according to the following protocol: current traces were obtained from voltage steps from +20 to +80 mV in 10 mV increments (holding potential was 0 mV); the sampling interval was 1.75 s.