Conflicting results have been reported on whether closed kinetic chain exercises (such as a leg press) may induce more balanced activation of vastus medialis (VM) and lateralis (VL) muscles compared with open kinetic chain exercise (such as pure knee extension). This study aimed to 1) compare between-vasti motor unit activity and 2) analyze the combined motor unit behavior from both muscles between open and closed kinetic chain exercises. Thirteen participants (four women, mean ± SD age: 27 ± 5 yr) performed isometric knee extension and leg press at 10, 30, 50, 70% of the maximum voluntary torque. High density surface EMG signals were recorded from the VM and VL and motor unit firings were automatically identified by convolutive blind source separation. We estimated the total synaptic input received by the two muscles by analyzing the difference in discharge rate from recruitment to target torque for motor units matched by recruitment threshold. When controlling for recruitment threshold and discharge rate at recruitment, the motor unit discharge rates were higher for knee extension compared with the leg press exercise at 50% [estimate = 1.2 pulses per second (pps), standard error (SE) = 0.3 pps, P = 0.0138] and 70% (estimate = 2.0 pps, SE = 0.3 pps, P = 0.0001) of maximal torque. However, no difference between the vasti muscles were detected in both exercises. The estimates of synaptic input to the muscles confirmed these results. In conclusion, the estimated synaptic input received by VM and VL was similar within and across exercises. However, both muscles had higher firing rates and estimated synaptic input at the highest torque levels during knee extension. Taken together, the results show that knee-extension is more suitable than leg-press exercise at increasing the concurrent activation of the vasti muscles.NEW & NOTEWORTHY There is a significant debate on whether open kinetic chain, single-joint knee extension exercise can influence the individual and combined activity of the vasti muscles compared with closed kinetic chain, multijoint leg press exercise. Here we show that attempting to change the contribution of either the vastus medialis or vastus lateralis via different forms of exercise does not seem to be a viable strategy. However, the adoption of open kinetic chain knee extension induces greater discharge rate and estimated synaptic input to both vasti muscles compared with the leg press.