Across studies, when presented with a variety of smoking cues, smokers and ex-smokers evidence distinct patterns of self-reported, physiological, and behavioral reactions. However, few studies have compared more than two different kinds of cues within the same experiment. Furthermore, despite the importance of examining the moderating effect of gender on smoking outcomes, few studies have examined gender differences in smoking cue reactivity. We examined the effect of eight distinct cue manipulations on heart rate, mean arterial pressure, smoking urges, and self-efficacy in a sample of 129 participants (50% female) who had recently quit smoking. Cue manipulations included (a) in vivo exposure, (b) an idiographically designed exposure of subjects' most recent relapse, (c) an idiographically designed exposure to subjects' highest risk situation, and (d) affectively valenced standardized scripts depicting situations generally associated with relapse. These manipulations were compared to a standard cognitive stressor (mental arithmetic) and to a resting baseline. Results revealed differences in the degree of reactivity to different manipulations, with in vivo cues producing the greatest changes. Gender differences in reactivity between the type of cues presented were found for mean arterial pressure, with standardized scripts producing greater changes for women. These findings have implications for understanding the reasons for differences in cue reactivity across manipulations and for gender differences in cue reactivity.