There is considerable interest in simple and effective methods to reduce sitting time and increase energy expenditure, and standing breaks have emerged as a realisable approach in individuals with obesity. The aim of the present study was to determine the extent to which energy expenditure in standing differs from sitting, and whether this energetic and metabolic-related responses are modified following a weight loss program in adolescents with obesity. After body composition assessment (DXA), cardiorespiratory and metabolic variables were continuously recorded (indirect calorimetry) during 10min while sitting and then during 5min standing posture before (n = 21; T1) and at the end of a multidisciplinary intervention (n = 17; T2) in adolescents with obesity. Before and after the intervention, energy expenditure and fat oxidation rates were significantly increased in standing compared with sitting. Weight loss did not change the relationship between sitting and standing energy expenditure. Sitting energy expenditure represented 1.0 and 1.1 Metabolic Equivalent of Task at T1 and T2, and increased to 1.1 and 1.2 during standing at T1 and T2, respectively. The percentage of change of android fat mass between T1 and T2 was positively associated with the percentage of change in energy expenditure from sitting to standing at T2. The vast majority of the adolescents with obesity significantly increased their energy expenditure between sitting and standing, both before and after a weight loss intervention. However, the standing posture did not allow breaking the sedentary threshold. Abdominal fat mass is associated with energic profile.