Acculturative stress is a salient contextual stressor for Latinx college students and may adversely impact their familial and psychological processes. Although much empirical literature has examined acculturative stress, family relations, and depressive symptoms among Latinx individuals, considerably less research has used longitudinal cross-lagged designs to clarify temporal relationships and directions of influence among the variables. To fill this research gap, the present study employed a longitudinal cross-lagged design to examine acculturative stress, family cohesion, family conflict, and depressive symptoms among Latinx college students (N = 173) with data collected 6 months apart between two time points. Cross-lagged findings indicated that higher levels of acculturative stress predicted an increase in family cultural conflict, but family cultural conflict did not longitudinally predict acculturative stress. Higher levels of family cultural conflict at Time 1 predicted higher levels of depressive symptoms at Time 2, but the reverse was not true. Finally, greater family cohesion predicted a decrease in family cultural conflict 6 months later, but family cultural conflict did not predict family cohesion across time. Implications for preventing Latinx college students' depressive symptoms are discussed with a threefold focus: (a) addressing family cultural conflict, (b) tackling acculturative stress to prevent family cultural conflict, and (c) increasing family cohesion to decrease family cultural conflict. Study limitations and future research directions are discussed. Public Significance Statement The present study with Latinx college students sought to clarify the temporal directions of influence among acculturative stress, family relations, and depressive symptoms. Findings illuminate that acculturative stress contributes to more family cultural conflict over time, and that family cultural conflict prospectively predicts higher levels of depressive symptoms. In contrast, family cohesion is a resource factor that can decrease family cultural conflict. Efforts in supporting Latinx college students' mental health should focus on addressing family cultural conflict, tackling acculturative stress, and increasing family cohesion.