Fig 5.

Representative Ca²⁺ fluorescence responses for Ca_V2.2 (A, C, E) and Na_V1.7 (B, D, F), transfected by calcium phosphate-, FuGENE-, and Lipofectamine 3000 measured in a FLIPR^TETRA. The KCl-evoked Ca²⁺ fluorescence responses of Ca_V2.2 over baseline for calcium phosphate-, FuGENE-, and Lipofectamine 3000 were 1.3 ± 0.0 (A), 1.7 ± 0.1 (C), and 1.5 ± 0.1 (E), respectively. The addition of 500 nM, and 16 nM CVID blocked the fluorescence response by 99.4 ± 1.8%, and 51.2 ± 3.3%, respectively, for calcium phosphate (A), 100.8 ± 0.9% and 50.1 ± 1.2%, respectively, for FuGENE (C), and 99.1 ± 1.0% and 49.5 ± 2.1%, respectively, for Lipofectamine 3000 (E). The veratridine-evoked Ca²⁺ fluorescence responses of Na_V1.7 over baseline for calcium phosphate-, FuGENE-, and Lipofectamine 3000-mediated transfetion were 0.9 ± 0.0 (B), 1.2 ± 0.1 (D), and 1.1 ± 0.1 (E), respectively. Addition of 1 μM, and 111 nM TTX blocked the fluorescence response by 98.9 ± 1.8% and 53.8 ± 1.6%, respectively, for calcium phosphate (B), 99.8 ± 0.4% and 54.2 ± 2.0%, respectively, for FuGENE (D), and 99.4 ± 1.1% and 51.2 ± 2.3%, respectively, for Lipofectamine 3000 (F). Data are represented as mean ± SEM (N = 3 independent experiments conducted in triplicate).