The anterior cingulate cortex (ACC) is critically involved not only in affective and anxiety processing, but also in error and conflict monitoring. To investigate how anxiety interacts with processing affective ambiguity, 15 anxious and 15 nonanxious individuals were scanned while performing a validated affective appraisal task, in which the fraction of faces of a particular affect or gender was parametrically controlled to provide various levels of ambiguity. The anxious group showed less ventral and greater dorsal ACC activation during ambiguous affective relative to ambiguous gender stimuli. For anxious individuals, dorsal ACC activation was related to a more biased response. Collectively, these data indicate that anxious individuals activate the dorsal and ventral components of the ACC differently during affective appraisal.