The primary purpose of this study was to determine the effects of a quadriceps/hip flexor Proprioceptive Neuromuscular Facilitation (PNF) stretch protocol on anterior pelvic tilt (APT), hamstring range of motion (ROM), and gluteal muscle activation. We also examined the relationship between hamstring ROM and gluteal activation. Eligible participants (N = 35; 24(± 4) years) completed an 8-minute stationary bike warm-up at a self-selected pace. Then participants completed measurements of standing sagittal assessment, Active Straight Leg Raise (ASLR), and Cook’s Hook test (CH) to measure APT, hamstring ROM, and gluteal activation, respectively. After a 4-minute rest, the measures were repeated. A quadriceps/hip flexor PNF stretch intervention (~ 4 minutes) was completed (10-6-30-second hold-relax method). The measures were repeated a third time. The study found 86% (right side), and 88% (left side) with APT initially improved their APT to normal after the PNF intervention. There was a significant bilateral improvement in hamstring ROM after the PNF intervention (p < 0.001). For gluteal activation, 30% (right side) and 45% (left side) improved from hamstring dominance to gluteal dominance after the PNF intervention. Lastly, those that were gluteal dominant (favorable) had significantly more hamstring ROM than those that were hamstring dominant at all time points (p < 0.001). These data support a hip flexors/quadriceps PNF stretch protocol were effective to immediately reduce APT, increase hamstring ROM and gluteal activation, thereby improving lumbo-pelvic posture.