Human emotions guide verbal and non-verbal behaviour during social encounters. During public performances, performers’ emotions can be affected directly by an audience’s attitude. The valence of the emotional state (positive or negative) of a broad range of animal species is known to be associated with a body and visual orientation laterality bias. Here, we evaluated the influence of an audience’s attitude on professional actors’ head orientation and gaze direction during two theatrical performances with controlled observers’ reactions (Hostile vs Friendly audience). First, our speech fluency analysis confirmed that an audience’s attitude influenced actors’ emotions. Second, we found that, whereas actors oriented more their head to the left (i.e. Right Hemisphere Bias) when the audience was hostile, they gazed more straight ahead at Friendly spectators. These results are in accordance with the Valence-Specific Hypothesis that proposes that processing stimuli with negative valences involves the right hemisphere (i.e. left eye) more than the left hemisphere.