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In the present study we investigated the mechanism inherent in the decision-making process in a single-trial with the game called Prisoner’s Dilemma (PD), in which players might simultaneously encounter different conditions that influence cooperation: namely, reciprocity, perceived control, and risk taking. We conducted 2 experiments to examine how, and to what extent, these conditions contribute to cooperation. By incorporating direct measurements of the potential conditions into a paradigm produced in previous studies, we were able to test the reliability of backward inference and to quantify the effect exerted by each condition. Results consistently revealed a complex cognitive process in single-trial PD: each condition had an influence on decision making, but none of them showed a dominating power to prompt cooperation independently; reciprocity served as a moderator.