Guilt is widely acknowledged as an exemplary social emotion that is unpleasant but has positive interpersonal consequences. Previous empirical research focuses largely on documenting the behavioral consequences of guilt; less is known about the psychophysiology of experiencing guilt. Here we designed an interactive paradigm and asked participants to play multiple rounds of a dot-estimation task with two partners. Failure in the task, either due to the participant or due to the partner, would cause electric shocks to the partner. In Experiment 1, we asked the participant to watch video clips depicting the partner's facial expressions while the partner was receiving pain stimulation. Eye movement recording showed that the participant fixated less on the partner's eyes but more on the nose region in the participant-caused pain (high guilt) condition than in the partner-caused pain (low guilt) condition, an indication of social avoidance. In Experiment 2, we asked the participant to either fixate on the eye (Eye Group) or the nose region (Nose group) of the partner and recorded their skin conductance during the viewing. We found that the Eye Group exhibited higher skin conductance response in the high guilt condition than in the low guilt condition and such a difference was absent for the Nose Group, indicating that the forced eye contact with the victim enhanced the emotional arousal of guilt. The life-like interactive paradigm is thus able to demonstrate the mutual dependence between eye contact and social emotions: eye contact both elicits and is regulated by emotional content in social interaction.