Driving behavior in expressway tunnels is more complicated than in common roadbed sections because of differences in illuminance, visual range, speed perception, and reaction time. To explore the setting method of exit advance guide signs in expressway tunnels and improve the effectiveness of drivers’ recognition of them, we propose 12 layout forms based on information quantification theory. In experiments, UC-win/Road was used to build a simulation scene, and the recognition reaction time of 12 element combinations of exit advance guide signs of different subjects was collected through an E-Prime simulation experiment. The loading effectiveness of the signs was analyzed based on the subjective workload and comprehensive evaluation scores of different subjects. The results are the following. The width of the exit advance guide sign layout in the tunnel is negatively correlated with the height of Chinese characters and the distance between the characters and the edge of the sign. The larger the height of Chinese characters and the distance between them and the edge of the sign, the smaller the maximum layout width of the sign. Considering the driver’s reaction time, subjective workload, sign recognition, amount of sign information, sign accuracy, and sign safety of 12 different information combinations, we suggest that exit advance guide signs in tunnels should be laid out as Chinese/English place name + distance + guide arrow.