Steady-state voltage-dependent activation and inactivation parameters of various rEag1 constructs.

Unless stated otherwise, the voltage-dependent gating property of rEag1 channels was determined from steady-state currents recorded with 3 mM external KCl. Reversal potentials (as determined from I–V curves) and steady-state current amplitudes were used to calculate channel conductances at different membrane potentials, which in turn were normalized to the maximum amplitude to obtain the relative Po. Alternatively, for rEag1-chimera P and N, isochronal tail currents (in 3 mM KCl) were normalized to the maximum amplitude to obtain corresponding relative Po values. Data points for non-inactivating and inactivating channels were fit with one and two Boltzmann functions, respectively (see Methods for more detail). Curve fitting parameters: V0.5a and V0.5i represent the half-maximal voltage for activation and inactivation, respectively; ka and ki the slope factor for activation and inactivation, respectively; and for inactivating channels only, b the non-inactivating relative Po at depolarized potentials. Data are shown as mean ± SEM (*, significantly different from rEag1-WT; t-test: pSteady-state voltage-dependent activation and inactivation parameters of various rEag1 constructs.除非另有说明，大鼠Eag1（rEag1）通道的电压依赖性门控特性，均通过3 mM胞外氯化钾（KCl）条件下记录的稳态电流进行测定。由电流-电压（I-V）曲线确定的逆转电位与稳态电流幅值，可用于计算不同膜电位下的通道电导，随后将电导归一化至最大幅值，以得到相对开放概率（relative Po）。针对rEag1嵌合体P与N（rEag1-chimera P and N），则采用3 mM KCl条件下的等时尾电流，将其归一化至最大幅值以获取对应的相对开放概率数值。非失活与失活通道的数据点，分别采用单组分玻尔兹曼（Boltzmann）函数与双组分玻尔兹曼函数进行拟合（详细方法参见材料与方法）。曲线拟合参数说明如下：V0.5a与V0.5i分别代表激活与失活的半最大电压；ka与ki分别为激活与失活过程的斜率因子；仅针对失活通道，参数b代表去极化电位下的非失活相对开放概率。所有数据均以平均值±标准误（SEM）表示（*表示与野生型rEag1（rEag1-WT）存在显著性差异；t检验：p<0.05）；本研究同时汇总了各类rEag1构建体的稳态电压依赖性激活与失活参数。