A pelvic position has been sought that optimizes abdominal muscle activation while diminishing hip flexor activation. Thus, the objective of the study was to investigate the effect of pelvic position and the Janda sit-up on trunk muscle activation. Sixteen male volunteers underwent electromyographic (EMG) testing of their abdominal and hip flexor muscles during a supine isometric double straight leg lift (DSLL) with the feet held approximately 5 cm above a board. The second exercise (Janda sit-up) was a sit-up action where participants simultaneously contracted the hamstrings and the abdominal musculature while holding an approximately 45 degrees angle at the knee. Root mean square surface electromyography was calculated for the Janda sit-up and DSLL under 3 pelvic positions: anterior, neutral, and posterior pelvic tilt. The selected muscles were the upper and lower rectus abdominis (URA, LRA), external obliques, lower abdominal stabilizers (LAS), rectus femoris, and biceps femoris. The Janda sit-up position demonstrated the highest URA and LRA activation and the lowest rectus femoris activation. The Janda sit-up and the posterior tilt were significantly greater (p < 0.01 and p < 0.05, respectively) than the anterior tilt for the URA and LRA muscles. Activation levels of the URA and LRA in neutral pelvis were significantly (p < 0.01 and p < 0.05, respectively) less than the Janda sit-up position, but not significantly different from the posterior tilt. No significant differences in EMG activity were found for the external obliques or LAS. No rectus femoris differences were found in the 3 pelvis positions. The results of this study indicate that pelvic position had a significant effect on the activation of selected trunk and hip muscles during isometric exercise, and the activation of the biceps femoris during the Janda sit-up reduced the activation of the rectus femoris while producing high levels of activation of the URA and LRA.