This study examined a pathway to heterosexual women's experience of anger and anxiety in response to lesbian interactions. Participants were 149 18-30 year old heterosexual female undergraduates (56% African American) from a southeastern United States university. Participants completed measures of female gender role beliefs, sexual prejudice, and state affect, viewed a video depicting relationship behavior between a female-female or male-female dyad, and again completed a measure of state affect. Results indicated that traditional beliefs about women were associated with higher levels of sexual prejudice toward lesbians. In turn, higher levels of sexual prejudice predicted increased anger (not anxiety) in response to the female-female, but not the male-female, dyad. Findings elucidate determinants of heterosexual women's anger, and potentially aggression, toward lesbians.