A left visual hemifield advantage for the processing of negative facial expressions has been demonstrated in a variety of studies. We tested whether the same effect is found for a neutral facial expression that had acquired a negative meaning through pairing with an aversive event. Startle reflex amplitudes, skin-conductance responses (SCR), and two verbal measures of affect (negative valence and arousal) were significantly increased after pairing, but no stimulation side by pairing interaction was observed. Functional magnetic resonance imaging (fMRI) revealed a significant increase of BOLD (blood oxygen level dependent) activity in the medial prefrontal cortex (MPFC), and the right frontal opercular region (RFOP). MPFC activity was correlated with psychophysiological and verbal emotional responses across subjects and, like these responses, was independent of the stimulation side. In contrast, RFOP activity was significantly stronger during left than during right hemifield stimulation but did not correlate with psychophysiological or verbal measures of negative affect. These results suggest that emotional responses to an aversive conditioned expressively neutral face are similar in both visual hemifields. MPFC activity seems to be closely linked to the strength of these responses.