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It is not uncommon for sedentary individuals to cite leg fatigue as the primary factor for test termination during a cardiopulmonary exercise test (CPET) on a cycle ergometer. The purpose of this study was to examine the effect of 2 weeks of lower body resistance training (RT) on cardiopulmonary capacity in sedentary middle-aged females. Additionally, the impact of RT on muscle strength was evaluated. Following familiarization, 28 women (18 exercise group, 10 control group) completed a maximal CPET on a cycle ergometer to determine peak oxygen uptake and leg extensor strength assessed using isokinetic dynamometry. Participants in the exercise group performed 2 weeks (6 sessions) of lower body RT, which comprised leg press, leg curl, and leg extension exercises. A 2-way repeated-measures ANOVA was used to evaluate the difference in changes of peak oxygen uptake and peak torque (PT). Peak oxygen uptake significantly improved from 22.2 ± 4.5 mL·kg−1·min−1 to 24.3 ± 4.4 mL·kg−1·min−1 (10.8%, p < 0.05) as well as PT from 83.1 ± 25.4 Nm to 89.0 ± 29.7 Nm (6.1%, p < 0.05) in the exercise group with no change in the control group. These findings provide initial evidence that 2 weeks of lower body RT prior to a CPET may be a helpful preconditioning strategy to achieve a more accurate peak oxygen uptake during testing, enhancing tolerability to a CPET by improving lower body strength.