Whole-cell currents were recorded using the patch-clamp technique at 27–28°C. hBM-MSC were trypsinized and seeded onto glass coverslips 30–60 min before assays. Bath solution contained (in mM): 140 NaCl, 4.5 KCl, 2 CaCl2, 1 MgCl2, and 10 [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] HEPES. The pipette solution contained (in mM): 140 CsCl, 2 ethylene glycol tetraacetic acid (EGTA), and 10 HEPES. pH was adjusted to 7.3–7.4. Pipettes were made from borosilicate glass capillaries (WPI, Inc; 1B150F-4), pulled in a horizontal micropipette puller (P-97, Sutter Instruments) and heat polished (MF-830, Narishige). Pipette resistance was 3–5 MΩ and seal resistance was > 1.5 GΩ. The cell mean capacitance was 27 ± 10 pF SEM (n = 3). Current recordings were acquired using an EPC-7 amplifier (HEKA), filtered at 1/5 of the acquisition rate and sampled with an A/D converter (NI-PCIe-6351; National Instruments). Acquisition software was developed in the LabView programming environment (National Instruments) by Dr. Patricio Orio (CINV, Valparaiso, Chile). Data analysis was performed with Clampfit 9 (Axon Instruments) software. Currents were activated by voltage ramp protocol from −100 to 200 mV (600 ms, every 2 s) with a holding potential (HP) of 0 mV. The voltage step protocol was also used with pulses between −100 and +150 mV in 10 mV increments with a duration of 45 ms. Currents were recorded before and after the addition of menthol 500 μM and BCTC 10 μM. Statistical analyses were performed using GraphPad Prism (One-way ANOVA).

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