DRG neurons were acutely dissociated from Sprague–Dawley (SD) rats as previously described [24 (link)]. DRG neurons with diameters of less than 10 mm were selected to study K+ channels, since small-sized DRG neurons tend to express Kv4.3 [8 (link)]. SD rats (Hunan SJA Laboratory Animal Co., Ltd., China) were used according to the guidelines of the National Institutes of Health for Care and Use of Laboratory Animals. The experiments were approved by the Animal Care and Use Committee of Hunan Normal University.
The human embryonic kidney 293 T cell line (HEK293T) was obtained from Shanghai Institute of Cell Biology (Chinese Academy of Sciences, China) and maintained in DMEM (Invitrogen, USA) supplemented with 10% heat-inactivated fetal calf serum (GibcoTM, USA), penicillin (100 U/ml, Sangon biotech, China), and streptomycin (100 μg/ml, Sangon biotech, China). Wild-type Kv4.3 or all of the mutants were transiently co-transfected into HEK293T cells, with eGFP using the LipfectamineTM2000 Reagent (Invitrogen, USA). The cells that had green fluorescence were selected to electrophysiological assays.
The human embryonic kidney 293 T cell line (HEK293T) was obtained from Shanghai Institute of Cell Biology (Chinese Academy of Sciences, China) and maintained in DMEM (Invitrogen, USA) supplemented with 10% heat-inactivated fetal calf serum (GibcoTM, USA), penicillin (100 U/ml, Sangon biotech, China), and streptomycin (100 μg/ml, Sangon biotech, China). Wild-type Kv4.3 or all of the mutants were transiently co-transfected into HEK293T cells, with eGFP using the LipfectamineTM2000 Reagent (Invitrogen, USA). The cells that had green fluorescence were selected to electrophysiological assays.
