The purpose of this work was to investigate the electromyographic (EMG) fatigue representations in muscles of male runners during run at different level of intensity. In this study, the EMG signals for the rectus femoris and biceps femoris (long head) were collected by bipolar electrodes from the left and right lower extremities. EMG measurements were recorded during the run on tartan athletic track. Four professional athletes had to run a 400 m distance with a different intensity. The first distance of 400 m took 90 s; the second, 70 s; the third, 60 s; and the last one was covered with a maximal velocity until exhaustion. Power spectral analysis of EMG signals was carried out to calculate MPF. The results of our study revealed the efforts of different intensity for each muscle individually. The effect of fatigue was observed only in the case of running with the highest velocity. The biggest changes in MPF were observed for BF (23.6%) and RF (19.5%) muscles of the left leg and then for BF (17.5%) and RF (12.5%) ones of the right leg. We supposed that those differences between the right and left legs were mainly due to the curve of the track where those muscles are differently loaded.