The ability to tell whether or not someone is lying has practical value in both the criminal justice system and in daily life. Prior research has demonstrated that training in acting can enhance empathy and theory of mind, which are two abilities that people draw on to determine whether or not someone is telling the truth. Is it possible, therefore, that acting experience can enhance lie detection capabilities? The present study addressed this question by recruiting participants with varying levels of acting experience to complete a lie detection task that involved assessing whether or not a student was lying about (not) cheating in a series of videotaped interviews. Participants also completed a microexpression identification task where they had to determine which of three negative emotional facial expressions (sad, angry, fearful) was briefly flashed on a face presented on the computer screen, as past research suggests that the ability to perceive microexpressions predicts lie detection performance. Finally, participants were given questionnaires designed to measure their levels of trait empathy and acting experience. This design allowed us to test the hypothesis that acting experience would lead to enhanced lie detection abilities, and examine whether this effect was mediated by an increase in empathy and/or microexpression identification skills. Results indicated that higher levels of empathy did marginally predict participants' accuracy in lie detection, and acting experience marginally predicted empathy levels. However, there was little evidence to link acting experience directly to increased empathy, microexpression identification skills, or lie detection. These findings suggest that further research may be necessary to fully understand the relationship between acting experience, empathy, and lie detection.