Context: Lesser hip muscle strength is commonly observed in females with patellofemoral pain (PFP) compared with females without PFP and is associated with poor subjective function and single-leg squat (SLS) biomechanics. Hip muscle weakness is theorized to be related to PFP, suggesting centrally mediated muscle inhibition may influence the observed weakness. The central activation ratio (CAR) is a common metric used to quantify muscle inhibition via burst superimposition. However, gluteal inhibition has yet to be evaluated using this approach in females with PFP. The study objectives are to (1) describe gluteal activation in the context of subjective function, hip strength, and squatting biomechanics and (2) examine the relationship of gluteal activation with subjective function and squatting biomechanics in females with PFP. Design: Cross-sectional. Methods: Seven females with PFP (age = 22.8 [3.6] y; mass = 69.4 [18.0] kg; height = 1.67 [0.05] m, duration of pain = 6–96 mo) completed this study. Subjective function was assessed with the Anterior Knee Pain Scale, while fear-avoidance beliefs were assessed with the Fear-Avoidance Belief Questionnaire physical activity and work subscales. Biomechanical function was assessed with peak hip and knee angles and moments in the sagittal and frontal planes during SLS. Gluteus medius (GMed) and gluteus maximus (GMax) activation were assessed with the CAR. Descriptive statistics were calculated, and relationships between variables were assessed with Spearman rho correlations. Results: The CAR of GMed and GMax was 90.5% (8.1%) and 84.0% (6.3%), respectively. Lesser GMed CAR was strongly associated with greater hip adduction during SLS (ρ = −.775, P = .02) and greater fear-avoidance beliefs—physical activity subscale (ρ = −.764, P = .018). Conclusion: We found a wide range in GMed and GMax activation across females with PFP. Lesser GMed activation was associated with greater hip adduction during SLS and fear of physical activity, suggesting that gluteal inhibition should be assessed in patients with PFP.