Interventional radiologists are at increased risk for musculoskeletal discomfort/disorders and this has been linked to the use of radiation personal protective equipment (rPPE). This study examined the effects of rPPE on the development of fatigue of the erector spinae and trapezius muscles. Surface electromyography (EMG) was used to capture muscle activity, and both time domain (average rectified value) and frequency domain (median frequency) measures were considered in the assessment of localized muscle fatigue. Sixteen participants performed a simulated surgical procedure requiring intermittent 30° flexed static trunk posture with and without rPPE on separate days. The results showed that the rPPE condition demonstrated significantly greater (p < 0.05) downward shift in median frequency in the left lumbar erector spinae and left lower thoracic erector spinae consistent with task-induced localized muscle fatigue. Ergonomic intervention strategies are discussed.