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Previous studies on emotion regulation have shown that cognitive reappraisal is a healthier and more effective emotion regulation strategy than expression suppression. Frontal electroencephalography (EEG) asymmetry reflects the difference between brain activation in left and right frontal areas and is widely viewed as an effective biomarker of emotional reactivity and regulation. In the present study, we investigated the asymmetry of the frontal EEG activity during the adolescents’ emotional regulation, and explored their relationship with the adolescents’ habitual use of emotional regulation strategies. Habitual use of cognitive reappraisal and expression suppression were measured by using the emotion regulation questionnaire (ERQ). EEG was recorded from 54 adolescents (24 boys, 30 girls, Mage=12.59), during the Reactivity and Regulation-Image Task. Result showed that, (1) in real-time emotion regulation tasks, there was a greater relative left frontal EEG activity under negative up-regulation conditions (compared to neutral up-regulation conditions); (2) habitual use of reappraisal was positively correlated with increased relative left frontal activity during negative emotion regulation. In contrast, no significant correlation was found between habitual use of suppression and frontal alpha asymmetry. The results provide neurological evidence that, for adolescents, the use of habitual emotion regulation strategies may affect the effect of real-time emotion regulation.