This study aims to advance the media effects debate concerning violent video games. Meta-analytic reviews reveal a small but noticeable association between playing violent video games and aggressive reactions. However, evidence for causal associations is still rare. In a novel, event-related functional magnetic resonance imaging study, 13 male research participants were observed playing a latest-generation violent video game. Each participant's game play was recorded and content analyzed on a frame-by-frame basis. Onscreen activities were coded as either "passive/dead, no a frame interactions"; "active/safe, no imminent danger/no violent interactions"; "active/potential danger occurs, violent interactions expected"; "active/under attack, some violent interactions"; and "active/fighting and killing, many violent interactions." Previous studies in neuroscience on aggressive thoughts and behaviors suggested that virtual violence would suppress affective areas of the anterior cingulate cortex (ACC) and the amygdala subsequent to activity variations at cognitive areas of the ACC. Comparison of game play activities with and without virtual violence in 11 participants confirmed the hypothesis. The rather large observed effects can be considered as caused by the virtual violence. We discuss the applicability of neuroscience methodology in media effects studies, with a special emphasis on the assumption of virtuality prevalent in video game play.