This study examined whether instructing mothers to apply emotion regulation strategies can change mothers’ perception and reactivity to infant crying in an experimental within-subject design. Perception of crying, skin conductance level (SCL), facial expressivity, and intended caregiving responses to cry sounds were measured in mothers (N = 101, M = 30.88 years) who received suppression, reappraisal, and no emotion regulation instructions. Reappraisal resulted in lower SCL during exposure to crying and a less negative perception of crying compared to the suppression condition. In contrast, suppression resulted in increased facial expressions of sadness compared to the control condition. Thus, simple instructions on how to reframe thoughts about crying can change mothers’ perception of and reactivity to crying.