The effect of gender stereotype activation on challenge/threat motivational states was examined. Male and female participants completed a difficult math test described as either gender-biased or gender-fair, while continuous cardiovascular data were recorded. During the math test, women in the gender-biased condition exhibited a threatened motivational state, whereas women in the gender-fair condition exhibited challenge. The cardiovascular pattern of data was reversed for men, with men exhibiting challenge when a gender bias was implied, but threat when it was not. Motivational implications of stereotype threat and psychophysiological measurement are discussed.