The purpose of this study was to examine EMG activities and VMO/VL ratio of the vastus medialis oblique, and vastus lateralis during step up exercise according to ankle and knee positions in soccer players with patellofemoral pain syndrome. Methods: Subject(patellofemoral pain syndrome, PFPS: n=8 and without PFPS, non PFPS; NPFPS: n=8) perfomed step up exercise at each knee and ankle position(knee flexion $30^{\circ}$ , $60^{\circ}$ , and $90^{\circ}$ , ankle internal rotation $30^{\circ}$ , neutral, and external rotation $30^{\circ}$ ) while EMG activity was collected. The EMG signals were expressed by the % maximal voluntary isometric Contraction(%MVIC) values. Statistical analysis consisted of two way repeated measures analysis of variance with post hoc analysis. Results: Main results were as follows: 1) EMG of VMO, and VL was tend to be lower in PFPS compared to NPFPS. 2) EMG of VMO and VL with knee flexrion $60^{\circ}$ was significantly higher the results with knee flexion $30^{\circ}$ , and $90^{\circ}$ . VMO and VL with ankle external rotation $30^{\circ}$ was significantly higher the results with internal rotation $30^{\circ}$ and neutral position. Conclusion: Considering the EMG activity was reduced due to the to the PFPS and that performing step up with knee flexion $60^{\circ}$ with ankle external rotation $30^{\circ}$ position may provide the most effective condition for patients with patellofemoral pain syndrome.