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This study investigated the effects of a novel, classroom-based emotion self-regulation program (TestEdge) on measures of test anxiety, socioemotional function, test performance, and heart rate variability (HRV) in high school students. The program teaches students how to self-generate a specific psychophysiological state – psychophysiological coherence – which has been shown to improve nervous system function, emotional stability, and cognitive performance. Implemented as part of a larger study investigating the population of tenth grade students in two California high schools (N = 980), the research reported here was conducted as a controlled pre- and postintervention laboratory experiment, using electrophysiological measures, on a random stratified sample of students from the intervention and control schools (N = 136). The Stroop color-word conflict test was used as the experiment’s stimulus to simulate the stress of taking a high-stakes test, while continuous HRV recordings were gathered. The postintervention electrophysiological results showed a pattern of improvement across all HRV measures, indicating that students who received the intervention program had learned how to better manage their emotions and to self-activate the psychophysiological coherence state under stressful conditions. Moreover, students with high test anxiety exhibited increased HRV and heart rhythm coherence even during a resting baseline condition (without conscious use of the program’s techniques), suggesting that they had internalized the benefits of the intervention. Consistent with these results, students exhibited reduced test anxiety and reduced negative affect after the intervention. Finally, there is suggestive evidence from a matched-pairs analysis that reduced test anxiety and increased psychophysiological coherence appear to be directly associated with improved test performance—a finding consistent with evidence from the larger study.