Influence of different load exercise to muscle activity during subsequent exercise with 75% of one repetition maximum (RM) load among trained and untrained individuals was verified. Resistance-trained men who were involved in resistance training (n = 16) and healthy young men who did not exercise regularly (n = 16) were recruited for this study. Each subject performed bench pressing with a narrow grip exercise using two different training set methods, the drop-set (DS) (3 sets × 2-10 repetitions with 95-75% of 1RM) and the reverse drop-set (RDS) (3 sets × 3-10 repetitions with 55-75% of 1RM). The mean concentric contraction power, root mean square (RMS) of electromyography (EMG), area under the oxygenated hemoglobin (Oxy-Hb) curve, and time constant for muscle oxygen consumption (TcVO_2mus) values of the triceps brachii were measured during and after the DS and RDS. The trained group demonstrated significantly higher mean muscle power (242.9 ± 39.6 W vs. 215.8 ± 31.7 W), RMS of EMG (86.4 ± 10.4 % vs. 68.3 ± 9.6 %), and area under the Oxy-Hb curve (38.6 ± 7.4 %• sec vs. 29.3 ± 5.8 %• sec) values during the DS than during the RDS (p < 0.05). However, in the untrained group none of the parameters differed significantly for both the DS and RDS. Furthermore, a negative correlation was detected between the area under the Oxy-Hb curve and muscle thickness (r = -0.51, p < 0.01). Long-term effects of DS on muscle strengthening and hypertrophy will be explored in further research.

• The DS induced greater motor unit activation and intramuscular hypoxia in people who have been regularly performing resistance exercises for more than one year than the RDS.

• No mechanical or metabolic differences were detected between the DS and RDS among the subjects who had not participated in regular resistance training.

• The thicker a person’s muscles are, the more resistant they are to the induction of acute intramuscular hypoxia during muscle contraction.