To determine whether electromyographic (EMG) onsets of vastus medialis obliquus (VMO) and vastus lateralis (VL) are altered in the presence of patellofemoral pain syndrome (PFPS) during the functional task of stair stepping. Cross-sectional. University laboratory. Thirty-three subjects with PFPS and 33 asymptomatic controls. Subjects ascended and descended a set of stairs-2 steps, each 20-cm high-at usual stair-stepping pace. EMG readings of VMO and VL taken on middle stair during step up (concentric contraction) and step down (eccentric contraction). Relative difference in onset of surface EMG activity of VMO compared with VL during a stair-stepping task. EMG onsets were determined by using a computer algorithm and were verified visually. In the PFPS population, the EMG onset of VL occurred before that of VMO in both the step up and step down phases of the stair-stepping task (p <.05). In contrast, no such differences occurred in the onsets of EMG activity of VMO and VL in either phase of the task for the control subjects. This finding supports the hypothesized relationship between changes in the timing of activity of the vastimuscles and PFPS. This finding provides theoretical rationale to support physiotherapy treatment commonly used in the management of PFPS.