Hypoxia negatively impacts aerobic exercise, but exercise testing in hypoxia has not been studied comprehensively. To determine the effects of simulated altitude on the gas exchange threshold (GET), respiratory compensation point (RCP), and maximal oxygen uptake (<(V)over dot>O(2)max), 24 participants (mean [SD]; 26 [4] years; 171.6 [9.7] cm; 69.2 [11.9] kg) acclimatized to mild altitude (MILD; similar to 1100 m) performed three cycling ramp-incremental exercise tests (with verification stages performed at 110% of peak power output (PPO)) in simulated altitudes of 0 m (sea level, SL), 1111 m (MILD), and 2222 m (moderate altitude, MOD), in a randomized order. There were significant effects of condition (i.e., fraction of inspired oxygen [FIO2]) for GET (p = 0.001), RCP (p < 0.001), <(V)over dot>O(2)max (p < 0.001), and PPO (p < 0.001). The <(V)over dot>O-2 corresponding to GET and RCP (mL center dot kg-1 center dot min-1) in MOD (24.1 [4.3]; 37.3 [5.1]) were significantly lower (p < 0.05) compared to SL (27.1 [4.4]; 41.8 [6.6]) and MILD (26.8 [5.7]; 40.7 [7.3]) but similar (p > 0.05) between SL and MILD. For each increase in simulated altitude, <(V)over dot>O(2)max (SL: 51.3 [7.4]; MILD: 50.0 [7.6]; MOD: 47.3 [7.1] mL center dot kg-1 center dot min-1) and PPO (SL: 332 [80]; MILD: 327 [78]; SL: 316 [76] W) decreased significantly (p < 0.05 for all comparisons). VO2max values from the verification stage were lower than those measured during the ramp-incremental test (p = 0.017). Overall, a mild simulated altitude had a significant effect on <(V)over dot>O(2)max and PPO but not GET and RCP, MOD decreased all four variables, and the inclusion of a verification stage had little effect on the determination of <(V)over dot>O(2)max in a group of young healthy adults regardless of the FIO2. Trial registration: Open Science Framework 10.17605/OSF.IO/ZTC9E.