Expectation of others’ cooperative behavior plays a core role in economic cooperation. However, the dynamic neural substrates of expectation of cooperation are little understood. To fully understand EOC behavior in more natural social interactions, the present study employed functional near-infrared spectroscopy hyperscanning to simultaneously measure pairs of participants’ brain activations in a modified prisoner’s dilemma game. The data analysis revealed the following results. Firstly, under the high incentive condition, team EOC behavior elicited higher interbrain synchrony in the right inferior frontal (...) gyrus than individual EOC behavior. Meanwhile, the IBS in the IFG could predict the relationship between empathy/agreeableness and EOC behavior, and this prediction role was modulated by social environmental cues. These results indicate the involvement of the human mirror neuron system in the EOC behavior and the different neural substrates between team EOC and individual EOC, which also conform with theory that social behavior was affected by internal and external factors. Secondly, female dyads exhibited a higher IBS value of cooperative expectation than male dyads in the team EOC than the individual EOC in the dorsal medial prefrontal cortex, while in the individual EOC stage, the coherence value of female dyads was significantly higher than that of male dyads under the low incentive reward condition in the rIFG. These sex effects thus provide presumptive evidence that females are more sensitive to environmental cues and also suggest that during economic social interaction, females’ EOC behavior depends on more social cognitive abilities. Overall, these results raise intriguing questions for future research on human cooperative behaviors. (shrink)