Fig. 4

Eomes deletion enhanced SSC regeneration after busulfan-induced testicular injury. (A) Representative images of testes from control and Eomes-cKO mice at PD90. Scale bar = 5 mm, n = 3. (B) Telephone (mg)-to-body weight (g) ratios of control and Eomes-cKO mice; ns, not significant; n = 3. (C) Hematoxylin and eosin (H&E)-stained testicular cross-sections from control and Eomes-cKO mice at PD90. Scale bars = 100 μm, n = 3. (D) Representative images of testes from control and Eomes-cKO mice at 0, 32 or 72 days postbusulfan treatment. Scale bar = 5 mm, n = 5. (E) Testes (mg)-to-body weight (g) ratios of control and Eomes-cKO mice at 0, 32 or 72 days postbusulfan treatment. *p value < 0.05, ns means not significant, n = 5. (F) Representative images of H&E-stained testicular cross-sections of control and Eomes-cKO mice at 32 or 72 days postbusulfan treatment. Asterisks indicate seminiferous tubules with complete recovery of spermatogenesis. Scale bars = 100 μm, n = 5. (G) Quantification of damaged seminiferous tubules in the testes of control and Eomes-cKO mice 32 or 72 days after busulfan treatment. A total of 1000 spermatogenic tubules were quantified. n = 5, *p value < 0.05. (H) Representative images of whole-mount immunostaining of LIN28A⁺ spermatogonia in the seminiferous tubules of busulfan-treated testes at different time points. Scale bars = 50 μm, n = 3. (I) Quantification of LIN28A⁺ spermatogonia per 500 Sertoli cells in testicular cross-sections of control and Eomes-cKO mice at 72 days postbusulfan treatment. At least 500 Sox9⁺ cells were counted for each sample. n = 5, *p value < 0.05