Substance use behaviors do not occur in isolation of one another and are not static over time. As adolescents age into early adulthood, there may be dynamic changes in their substance use behaviors, and these changes may be influenced by family and school factors. The current study uses Latent Transition Analysis to examine these changes by measuring transitions among different substance use profiles based on past 30-day alcohol, tobacco and marijuana use, and by estimating associations with demographic, family and school factors. Data were from youth (n = 850; 80% African American, 17% white, 3% mixed race, 50% female and 50% male) in grade 10 (Time 1), with 24- (Time 2) and 48-month (Time 3) follow-ups. Substance use profiles included Non-users (54%), Alcohol and Marijuana Users (20%), and Alcohol, Tobacco and Marijuana Users (26%). There were considerable transitions among profiles from Time 1 to Time 2, and fewer transitions from Time 2 to Time 3. At Time 1, African American race and positive school attitudes were negatively associated with being an Alcohol and Marijuana User, and being an Alcohol, Tobacco and Marijuana User. Family conflict, parental school involvement, female gender and African American race were associated with transitions among substance use profiles. Implications are discussed for a better understanding of transitions in substance use profiles, and for promoting maintenance of non-use and transitions from substance using profiles to non-use.