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Abstract Background Toddlerhood (2–3 years) is a crucial period for the development of physical activity, sleep, and emotional self-regulation skills. Although there is growing evidence of positive associations between physical activity, sleep, and emotional self-regulation in school-aged children, the associations in toddlers remain unclear. Thus, this study aimed to examine the association between physical activity, sleep, and emotional self-regulation in toddlers. Methods Baseline data from 1350 toddlers (2.2 ± 0.33 year) from the Let’s Grow randomised controlled trial were used. Toddlers’ total physical activity (TPA) and moderate- to vigorous-intensity physical activity (MVPA) were assessed via 4 + days of accelerometry and a 3-item parent-report scale. Parent-reported total sleep quantity was calculated using the sum of average night-time sleep and daytime nap durations. Sleep behaviour data including bedtime routine, bedtime resistance, sleep onset-delay, sleep duration, and night waking were collected using relevant subsections from the Child Sleep Habits Questionnaire (CSHQ) and Brief Infant Sleep Questionnaire revised (BISQ-R). A 4-item parent-report scale adapted from the Fast Track Project Child Behaviour Questionnaire was used to assess toddlers’ emotional self-regulation. Linear regression models were used, adjusting for child age, sex, and parental education. Results Accelerometer-derived TPA, MVPA and parent-reported TPA were not associated with emotional self-regulation. Higher parent-reported MVPA (B = -0.01 CI95 -0.03, -0.003) was associated with poorer emotional self-regulation. Higher sleep duration was associated with better emotional self-regulation (B = 0.06 CI95 0.04, 0.08). The five sleep behaviours assessed were also positively associated with emotional self-regulation (all p |