Cocaine block of human cardiac (hH1) and rat skeletal (mu1) muscle sodium channels was examined under whole-cell voltage clamp in transiently transfected HEK293t cells. Low affinity block of resting mu1 and hH1 channels at -180 mV was the same, and high affinity block of inactivated channels at -70 mV was the same. Cocaine block of hH1 channels was greater than block of mu1 channels at voltages between -120 mV and -90 mV, suggesting that greater steady-state inactivation of hH1 channels in this voltage range makes them more susceptible to cocaine block. We induced shifts in the voltage dependence of steady-state inactivation at mu1 and hH1 channels by constructing mu1/hH1 channel chimeras or by coexpressing the wild-type channels with the rat brain beta1 subunit. In contrast to several previous reports, coexpression of the rat brain beta1 subunit with mu1 or hH1 produced large positive shifts in steady-state inactivation. Shifts in the voltage dependence of steady-state inactivation elicited linear shifts in steady-state cocaine block, yet these manipulations did not affect the cocaine affinity of resting or inactivated channels. These data, as well as simulations used to predict block, indicate that state-dependent cocaine block depends on both steady-state inactivation and channel activation, although inactivation appears to have the predominant role.