*4.2. Cell Culture and Transfection*

Chinese hamster ovary (CHO) cells, purchased from Evrogen company (Evrogen, Moscow, Russia), were cultured in a humidified atmosphere of 5% CO2 at 37 ◦C. Standard culture conditions were used for cell maintenance (Dulbecco's modified Eagle's medium (DMEM), 10% fetal bovine serum, 5% gentamicin). Transfection of plasmid encoding rat ASIC3 subunit was done using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer s protocol. We received expression vectors encoding rat ASIC3 as a gift from A. Staruschenko [59]. Those vectors were described in Reference [60]. Cells were transfected with 0.5 mg rASIC3 cDNA + 0.5 mg eGFP per 35-mm dish to achieve the expression of homomeric channels.
