Purpose The purpose of this study is to investigate the effects of grip width and hand orientation on muscle activities of upper body during the lat pull-down. Methods Eight healthy men performed the 6 grip variations (3 grip width × 2 hand orientation) using an experimentally determined load of 70% of 1 repetition maximum. Five trials were analyzed for each grip type. Participants maintained a cadence of 2-second concentric and 2-second eccentric phases. The grip widths were normalized for each individual by using their biacromial diameter (100%), and then set the grip width 50% of biacromial diameter and 150% of biacromial diameter. Surface EMG of the latissimus dorsi, pectoralis major, infraspinatus, biceps brachii, triceps brachii, rectus abdominis, erector spinae and middle trapezius was recorded, and the root mean square of the EMG was normalized, using a maximum isometric voluntary contraction. Results Latissimus dorsi showed higher muscle activities in 100% grip width than those of 50% grip width. Pectoralis major and rectus abdominis showed higher muscle activities in 50% and 100% grip width than those of 150% grip width. Middle trapezius showed higher muscle activities in 150% grip width than those of 50% grip width. Conclusions Two-way repeated measures ANOVA for each muscle revealed that latissimus dorsi and middle trapezius (the posterior muscles of trunk) showed higher muscle activities in wide grip, on the other hand, pectoralis major and rectus abdominis (the posterior muscles of trunk) showed higher muscle activities in narrow grip.