In this study, we examined longitudinal, person-centered trajectories of acculturation, internalizing symptoms, and self-esteem in 349 Latino adolescents. We compared acculturation measures (time in the US, culture-of-origin involvement, US cultural involvement, for both parents and adolescents); acculturation stressors (perceived discrimination, acculturation conflicts); and family dynamics (parent–adolescent conflict, familism). Results indicated that, over time, Latino adolescents’ internalizing problems decreased and their self-esteem increased. However, we showed that increased length of time living in the US was significantly related to lower self-esteem among adolescents. Parent–adolescent conflict was a strong risk factor, which not only directly heightened internalizing symptoms and lowered self-esteem, but also mediated the effects of acculturation conflicts and perceived discrimination on these outcomes. Our findings revealed familism as a cultural asset associated with fewer internalizing symptoms and higher self-esteem. Internalizing symptoms were also minimized by the adolescent’s involvement in the US culture whereas bicultural adolescents with high culture-of-origin involvement reported higher self-esteem. We discussed the limitations and implications of this study for future research and practice.