Eco-driving is a driving mode through driver behaviors mainly about accelerator works, such as smooth acceleration at start, preservation of steady speed, and active uses of engine brake. In this study, in virtue of a universal driving simulator (DS), driving experiments by subjects were conducted in the modes of normal driving, eco-driving, and eco-driving with eco-indicator. In the experiments, surface electromyography (sEMG) of leg muscles of driver was measured to clarify muscle fatigues in the different driving modes, and effect of mental workload was also analyzed from a subjectivity evaluation of NASA-TLX. The studies validated that the fuel economies can be improved by the eco-driving mode; however, the eco-driving requires more muscular activities than the normal driving. But the experimental results suggested that the muscular activities can be decelerated by using the co-indicator.