Previous studies have demonstrated that the neural basis of deception involves a network of regions including the medial frontal cortex (MFC), superior temporal sulcus (STS), temporo-parietal junction (TPJ), etc. However, to test the actual activity of the brain in the act of deceptive practice itself, existing studies have mainly adopted paradigms of passive deception, where participants are told to lie in certain conditions, and have focused on intra-brain mechanisms in single participants. In order to examine the neural substrates underlying more natural, spontaneous deception in real social interactions, the present study employed a functional near-infrared spectroscopy (fNIRS) hyperscanning technique to simultaneously measure pairs of participants’ fronto-temporal activations in a two-person gambling card-game. We demonstrated higher TPJ activation in deceptive compared to honest acts. Analysis of participants’ inter-brain correlation further revealed that the STS is uniquely involved in deception but not in honesty, especially in females. These results suggest that the STS may play a critical role in spontaneous deception due to mentalizing requirements relating to modulating opponents’ thoughts. To our knowledge, this study was the first to investigate such inter-brain correlates of deception in real face-to-face interactions, and thus is hoped to provide a new path for future complex social behavior research.