There is little information on behavioral and physical effects of lesson horses being used multiple times a day or ridden by riders of varying levels of skill, leaving lesson program managers with limited information to support horse management and welfare decisions. This study used video analysis to evaluate whether horses exhibited different limb kinematics or patterns of behavior under riders with differing levels of experience, factors that could impact physical effort by the horse. Riders (n = 8) were sorted by skill level (four beginner and four advanced), and horses (n = 8) were sorted by sensitivity level (four reactive and four nonreactive). Then pairs of horses (one reactive and one nonreactive in each pair) and pairs of riders (one beginner and advanced in each pair) were created. The pairs were then used in a repeated 2 × 2 Latin square design. Data were collected at the posting trot, using an English saddle, during the stance phase of single fore and hind footfalls. Multivariate analysis of variance of the kinematic variables revealed no overall trends across the kinematic variables, with only a small number of joints showing near-significant effects. Behaviors were quantified based on a study-specific ethogram and willingness scale, but no differences related to rider skill level or horse sensitivity were identified. Although our data suggest no differences in horse kinematics or behavior between beginner and advanced rider groups of horses ridden at the trot, further trials would be required to test for effects during other portions of the stride cycle, other gaits, or longer durations of locomotion.