Previous studies exploring the watching eyes effect with experimental games predominantly employed settings without real-interaction. However, the use of real-interaction setting is crucial for investigating replication and robustness of the watching eyes effect on prosocial behaviours, as prosocial behaviours primarily occur interpersonal interaction. Consequently, the present study comprised five experiments to examine the watching eyes effect on prosocial behaviours using oTree platform, which facilitates real interactions among participants. Furthermore, the current study compares the watching eyes effect across four principal types of prosocial behaviours - namely the Dictator Game, Ultimatum Game, Trust Game, and Prisoner's Dilemma - in real interaction settings. Our results indicated that in real interaction settings, this effect only occurs in the altruistic behaviour of dictator games. In both Experiments 1a (dictator game in laboratory) and 1b (dictator game online), dictators in the Eye condition distributed more amount to recipient and received a lower payoff than those in the Flower condition. However, the watching eyes effect in ultimatum game (Experiment 2), trust game (Experiment 3) and prisoner's dilemma (Experiment 4) was not evident. No significant difference were observed in target behaviours between the Eye and the Flower conditions in Experiments 2 (ultimatum game), 3 (trust game), and 4 (prisoner's dilemma). Our study was an attempt to examine the watching eyes effect in a series of prosocial behaviours with real interaction in one integrated platform, and compare this effect with the same dictator game online and in the laboratory. This study suggests that eye cues can potentially enhance prosocial behaviors in specific contexts such as blood donation and nature conservation, though further replication studies are required to confirm these findings.