BACKGROUND: The current study examined the acute effects of a lower body resistance training (RT) session on physiological and thermoregulatory measures during a sub-maximal running protocol in the heat in heat-acclimatized men. Ten resistance-untrained men (age 27.4 ± 4.1 years; height 1.78 ± 0.06 m; body mass 76.8 ± 9.9 kg; peak oxygen uptake 48.2 ± 7.0 mL kg-1 min-1) undertook a high-intensity RT session at six-repetition maximum. Indirect muscle damage markers (i.e., creatine kinase [CK], delayed-onset muscle soreness [DOMS], and countermovement jump [CMJ]) were collected prior to, immediately post and 24 and 48 h after the RT session. The sub-maximal running protocol was performed at 70% of the ventilatory threshold, which was conducted prior to and 24 and 48 h following the RT session to obtain physiological and thermoregulatory measures. RESULTS: The RT session exhibited significant increases in DOMS (p < 0.05; effect size [ES]: 1.41-10.53), whilst reduced CMJ (p < 0.05; ES: - 0.79-1.41) for 48 h post-exercise. There were no differences in CK (p > 0.05), although increased with moderate to large ES (0.71-1.12) for 48 h post-exercise. The physiological cost of running was increased for up to 48 h post-exercise (p < 0.05) with moderate to large ES (0.50-0.84), although no differences were shown in thermoregulatory measures (p > 0.05) with small ES (0.33). CONCLUSION: These results demonstrate that a RT session impairs sub-maximal running performance for several days post-exercise, although thermoregulatory measures are unperturbed despite elevated muscle damage indicators in heat-acclimatized, resistance untrained men. Accordingly, whilst a RT session may not increase susceptibility to heat-related injuries in heat-acclimatized men during sub-maximal running in the heat, endurance sessions should be undertaken with caution for at least 48 h post-exercise following the initial RT session in resistance untrained men.