Longo, AR, Silva-Batista, C, Pedroso, K, de Salles Painelli, V, Lasevicius, T, Schoenfeld, BJ, Aihara, AY, de Almeida Peres, B, Tricoli, V, and Teixeira, EL. Volume load rather than resting interval influences muscle hypertrophy during high-intensity resistance training. J Strength Cond Res XX(X): 000–000, 2020—Interset rest interval has been proposed as an important variable for inducing muscle mass and strength increases during resistance training. However, its influence remains unclear, especially when protocols with differing intervals have equalized volume. We aimed to compare the effects of long (LI) vs. short rest interval (SI) on muscle strength (one repetition maximum [1RM]) and quadriceps cross-sectional area (QCSA), with or without equalized volume load (VL). Twenty-eight subjects trained twice a week for 10 weeks. Each subject's leg was allocated to 1 of 4 unilateral knee extension protocols: LI, SI, SI with VL -matched by LI (VLI-SI), and LI with VL-matched by SI (VSI-LI). A 3-minute rest interval was afforded in LI and VSI-LI protocols, while SI and VLI-SI employed a 1-minute interval. All subjects trained with a load corresponding to 80% 1RM. One repetition maximum and QCSA were measured before and after training. All protocols significantly increased 1RM values in post-training (p < 0.0001; LI: 27.6%, effect size [ES] = 0.90; VLI-SI: 31.1%, ES = 1.00; SI: 26.5%, ES = 1.11; and VSI-LI: 31.2%, ES = 1.28), with no significant differences between protocols. Quadriceps cross-sectional area increased significantly for all protocols in post-training (p < 0.0001). However, absolute changes in QCSA were significantly greater in LI and VLI-SI (13.1%, ES: 0.66 and 12.9%, ES: 0.63) than SI and VSI-LI (6.8%, ES: 0.38 and 6.6%, ES: 0.37) (both comparisons, p < 0.05). These data suggest that maintenance of high loads is more important for strength increases, while a greater VL plays a primary role for hypertrophy, regardless of interset rest interval.