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This study investigated the acute effects of inter-set static stretching (ISS) during resistance exercise (RE) on the subsequent neuromuscular and metabolic responses. Twelve resistance-trained men performed three different knee extension RE protocols comprised of seven sets of 10 repetitions in a counterbalanced fashion. The three protocols were: 1) ISS (subjects performed 25 sec of quadriceps stretching between sets during 40 sec rest interval); 2) control (CON, subject passively rested between sets for 40 sec); 3) traditional (TRA, subject passively rested between sets for 120 sec). Total work was lower (p < 0.05) in ISS than CON and TRA (p <0.05). The fatigue index was greater (p < 0.05) in ISS compared with CON and TRA. ISS also resulted in lower (p < 0.05) electromyography (EMG) amplitude during the 6th and 7th sets compared with TRA. Additionally, EMG frequency was lower (p < 0.05) from the 3rd to 5th sets during ISS compared to CON, and from the 3rd to 7th sets compared to TRA. Muscle swelling and blood lactate similarly increased (p > 0.05) in response to all protocols. These results indicate that ISS negatively impacts neuromuscular performance, and does not increase the metabolic stress compared to passive rest intervals.