Objective. Mothers’ affective responses to infant laughing are essential in parent-child interaction. This experimental study examined whether instructing mothers to employ emotion regulation strategies can change their self-reported, physiological, and facial expressive responses to infant laughing. Design. Using a within-subjects design, mothers (N = 100, age M = 30.87 years) were exposed to infant laughing sounds while receiving enhancement, suppression, and no emotion regulation instructions. Positive affect, perception of laughing, intended sensitive and insensitive caregiving responses, skin conductance level, and facial expressions in response to infant laughing were measured. Results. Enhancement resulted in increased positive affect, a more positive perception of the laugh, more intended sensitive caregiving responses, and, compared to suppression, fewer intended insensitive caregiving responses. Moreover, enhancement resulted in lower sad and, compared to suppression, higher happy facial expressi-vity. In contrast, suppression resulted in a less positive perception. Enhancement did not affect skin conductance level. Conclusions. Enhancement can have beneficial effects on mothers’ self-reported and facial expressive responses to infant laughing in an experimental setting. Enhancement instructions may be used to increase mothers’ positive feelings in response to infant laughing and to promote sensitive caregiving behaviors and positive facial expressions, which may benefit affective mother-child interchanges.