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Previous studies have revealed that stereotyping processes involving negative affective content (e.g., antipathy) can lead to a significant increase in insula activity. However, whether the insula is sensitive to stereotype inconsistency or plays a crucial role in stereotype regulation remains unclear. To help fill this gap, 21 young adults were presented with a modified single-category implicit association test (SC-IAT) that assessed their stereotypes about rural migrant workers. In a within-subjects design, participants completed separate blocks of compatible and incompatible trials, each of which consisted of stereotype-consistent or stereotype-inconsistent trait labels that had a positive or a negative valence. Functional MRI was used to identify specific brain regions associated with negative and positive stereotyping. The behavioral results revealed a typical stereotype regulation effect in which participants responded slower to stereotype-inconsistent condition than stereotype-consistent condition, although such effect was significantly modulated by IAT compatibility, rather than by emotional valence. MRI results revealed that activity in the right insula was significantly sensitive to stereotype regulation processes in negative incompatible tasks, whereas such effect was marginally significant in positive incompatible tasks. Moreover, psychophysiological interaction analyses indicated complex functional connectivity among the right insula and cognitive control regions [e.g., ventrolateral prefrontal cortex (vlPFC) and dorsal anterior cingulate cortex (dACC)], social mentalizing regions [medial prefrontal cortex (mPFC)], and motivation regions (putamen) in the condition where negative stereotypes were violated. Taken together, these findings demonstrate that the right insula serves as a crucial node in regulating implicit stereotyping, particularly in negative stereotyping tasks. [ABSTRACT FROM AUTHOR]