This study examined whether the efficacy of keepin' it REAL, a model program for substance use prevention in schools, was moderated by gender, ethnicity, and acculturation. Gender differences in program efficacy may arise through boys' higher risk of drug use, inadequate attention to girls' developmental issues, or cultural factors like polarized gender expectations. Data came from a randomized trial in 35 Phoenix, Arizona, middle schools involving 4,622 mostly Latino 7th graders. Using multi-level mixed models and multiple imputation missing techniques, results for the total sample showed no gender differences in program effects on recent substance use, but the program was more effective in fostering boys' than girls' anti-drug norms. Subgroup analyses demonstrated several more beneficial program effects for boys than girls (less alcohol and cigarette use and stronger anti-drug norms), but only among less acculturated Latinos. There were no gender differences in program effects among more acculturated Latinos, nor among non-Latino whites.