The rating of perceived exertion (RPE) scale has been found to reflect physiological responses, and this study aimed to assess the validity of using the Borg CR-10 scale and velocity loss to evaluate muscle fatigue quantified by surface electromyography during back squat (BS) exercise. A total of 15 collegiate male athletes underwent three non-explosive BS tasks comprising low, medium, and high volumes at 65% of their one-repetition maximum. RPEs, spectral fatigue index (SFI), and velocity loss during BS exercise were assessed throughout the trials. Significant differences in overall RPE (p < 0.001) and average SFI (p < 0.05) were observed between the conditions, whereas no significant difference was observed in average velocity loss. Significant increases in RPE and SFI (p < 0.001) were observed within the exercise process, whereas a significant increase in velocity loss was not observed. Correlation analyses indicated a significant correlation between RPE and SFI obtained during exercise (r = 0.573, p < 0.001). However, no significant correlation was observed between velocity loss and SFI. These results demonstrated that RPE could be used as a muscle fatigue predictor in BS exercise, but that velocity loss may not reflect muscle fatigue correctly when participants cannot and/or are not required to perform BS explosively. Furthermore, practitioners should not use velocity loss as a muscle fatigue indicator in some resistance exercise situations, such as rehabilitation, beginner, and hypertrophy programs.