The purpose of the study was to compare force output and muscle activity of leg and trunk muscles in isometric squats executed on stable surface (i.e., floor), power board, BOSU ball, and balance cone. Fifteen healthy men (23.3 ± 2.7 years, mass: 80.5 ± 8.5 kg, height: 1.81 ± 0.09 m) volunteered. The force output and electromyographic (EMG) activities of the rectus femoris, vastus medialis, vastus lateralis, biceps femoris, soleus, rectus abdominis, oblique external, and erector spinae were assessed. The order of the surfaces was randomized. One familiarization session was executed before the experimental test. Compared with stable surface (749 ± 222 N), the force output using power board was similar (-7%, p = 0.320) but lower for BOSU ball (-19%, p = 0.003) and balance cone (-24%, p ≤ 0.001). The force output using BOSU ball and balance cone was approximately 13% (p = 0.037) and approximately 18% (p = 0.001) less than the power board. There were similar EMG activities between the surfaces in all muscles except for rectus femoris, in which stable squat provided greater EMG activity than did the other exercises (p = 0.004-0.030). Lower EMG activity was observed in the rectus femoris using balance cone compared with the BOSU ball (p = 0.030). In conclusion, increasing the instability of the surface during maximum effort isometric squats usually maintains the muscle activity of lower-limb and superficial trunk muscles although the force output is reduced. This suggests that unstable surfaces in the squat may be beneficial in rehabilitation and as a part of periodized training programs, because similar muscle activity can be achieved with reduced loads.