In recent years, there has been remarkable change in societal acceptance of lesbians and gay men. This meta-analysis explored whether this positive shift has reduced the gender difference in these attitudes. We tested Kite and Whitley’s (1996) gender belief system model and replicated their finding that heterosexual men held more negative attitudes toward gay people (g. = 0.37, k = 245, N = 98,295), gay behavior (g. = 0.22, k = 68, N = 98,734), and gay civil rights (g. = 0.16, k = 80, N = 89,187). We also found that heterosexual men were more likely to report engaging in anti-gay behavior (g. = 0.53, k = 7, N = 2,509), endorsing gay stereotypes (g. = 0.17, k = 17, N = 6,936), and wanting to avoid contact with gay men (g. = 0.49, k = 7, N = 2,178). However, heterosexual women reported a stronger desire to avoid contact with lesbians than did heterosexual men (g. = -0.36, k = 5, N = 1,339). For the most part, gender differences remained stable over time; the exceptions were attitudes toward same-gender sexual behavior and attitudes toward lesbian and gay civil rights, which showed small increases. Gender differences in anti-gay prejudice were mediated by gender differences in traditional gender-role beliefs. Moderator variables included participant group, type of measure, and target group.