<jats:p>Background: This study tested muscle activity (EMG) and perception of effort in eccentric (ECC) and concentric (CON) cycling before and after four sessions of both. Methods: Twelve volunteers naïve to ECC cycling attended the laboratory six times. On day 1, they performed a CON cycling peak power output (PPO) test. They then carried-out four sessions comprising two sets of 1 to 1.5-min cycling bouts at 5 intensities (30, 45, 60, 75, and 90% PPO) in ECC and CON cycling. On day 2 and day 6 (two weeks apart), EMG root mean square of the vastus lateralis (VL), rectus femoris (RF), biceps femoris (BF), and soleus (SOL) muscles, was averaged from 15 to 30 s within each 1-min bout and perception of effort was asked after 45 s. Results: Before the four cycling sessions, while VL EMG was lower in ECC than CON cycling, most variables were not different. Afterwards, ECC cycling exhibited lower RF EMG at 75 and 90% PPO (all p < 0.02), lower VL and BF EMG at all exercise intensities (all p < 0.02), and inferior SOL EMG (all p < 0.04) except at 45% PPO (p = 0.07). Perception of effort was lower in ECC cycling at all exercise intensities (all p < 0.03) but 60% PPO (p = 0.11). Conclusions: After four short sessions of ECC cycling, the activity of four leg muscles and perception of effort became lower in ECC than in CON cycling at most of five power outputs, while they were similar before.</jats:p>