Patch clamp recordings
All patch-clamp recordings in two different configurations of
macroscopic whole-cell and microscopic inside-out patch clamp recordings
were conducted after 12-24 h transfection with an EPC10 amplifier
powered by PatchMaster software (HEKA) at room temperature (23-25℃). The
borosilicate glass pipettes were pulled using a DMZ universal electrode
puller (Zeitz-Instruments GmbH) with the resistances of 3-5 MΩ (whole
cell) or 5-7 MΩ (inside-out) when
filled with a solution containing (in mM): (1) 150 NaCl, 2 EGTA, 10
HEPES, 1 InsP6 (pH 7.2, adjusted with NaOH) for humanTRPA1 (inside-out);
(2) 130 NaCl, 0.2 EGTA, 3 HEPES (pH 7.2, adjusted with NaOH) for others.
External solution was prepared ditto to internal solution. Currents were
recorded at a potential ramp from -100mV to +100 mV (whole-cell) or a
potential of +80 mV (inside-out).