The purpose of this cross-sectional study was to analyze changes in normalized surface electromyography (sEMG) signals for the gastrocnemius medialis, biceps femoris, gluteus maximus, tibialis anterior, and vastus lateralis muscles occurring during a 400 m indoor sprint between subsequent curved sections of the track. Ten well-trained female sprinters (age: 21 ± 4 years; body mass: 47 ± 5 kg; body height: 161 ± 7 cm; 400 m personal best: 52.4 ± 1.1 s) performed an all-out 400 m indoor sprint. Normalized sEMG signals were recorded bilaterally from the selected lower limb muscles. The two-way ANOVA (curve × side) revealed no statistically significant interaction. However, the main effect analysis showed that normalized sEMG signals significantly increased in subsequent curves run for all the studied muscles: gastrocnemius medialis (p = 0.003), biceps femoris (p < 0.0001), gluteus maximus (p = 0.044), tibialis anterior (p = 0.001), and vastus lateralis (p = 0.023), but differences between limbs were significant only for the gastrocnemius medialis (p = 0.012). The results suggest that the normalized sEMG signals for the lower limb muscles increased in successive curves during the 400 m indoor sprint. Moreover, the gastrocnemius medialis of the inner leg is highly activated while running curves; therefore, it should be properly prepared for high demands, and attention should be paid to the possibility of the occurrence of a negative adaptation, such as asymmetries.