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This study investigates how emotion influences learning performance (recognition, recall, and transfer test scores) and self-regulation in multimedia learning on health. Based on the Limited Capacity Model of Motivated Mediated Message Processing (LC4MP), this study uses a 3 (valence: positive, negative, coactive) × 2 (arousal: calm, arousing) experimental design. The findings will contribute to the understandings of the effects of emotion on multimedia learning and provide beneficial implications for health education.