This study aimed to monitor the oxygenation and blood supply in three quadriceps muscles [the vastus lateralis (VL), vastus medialis (VM), and rectus femoris (RF)] during squatting exercise to exhaustion. Eighteen young resistance-trained males performed five sets of 15 back squats in a Smith machine, with two warm-up sets [at 14% and 45% of the 15-repetition maximum (15RM)] and three main sets at 100% of the 15RM. Three near-infrared spectroscopy devices were attached to the VL, VM, and RF to record the muscle oxygen saturation (SmO2) and total hemoglobin (tHb, an index of muscle blood supply). The blood lactate concentration was measured after each set with a portable analyzer. The SmO2 and tHb data were analyzed by repeated-measures two-way ANOVA (muscle × set). Lactate data were analyzed by repeated-measures one-way ANOVA. The statistical significance was set at α = 0.05. The SmO2 dropped during each set (hitting zero in many instances) and was reinstated during recovery. The three main sets caused severe deoxygenation in the VL and VM, as opposed to moderate deoxygenation in the RF. From one set to the next, the initial value and the drop in the SmO2 increased, whereas the final SmO2 value decreased. The tHb increased in the VL, did not change considerably in the VM, and decreased in the RF during each set. The blood lactate concentration increased gradually from one set to the next, reaching about 10 mmol/L. These findings show pronounced differences in the physiological and metabolic responses of three quadriceps muscles to squatting exercise, thus highlighting the importance of studying such responses at multiple sites.