Fig. 6: Effects of GSH and Fer-1 on 8-month-old S8/Gss^−/− mice.

A Statistical results of litter size of mice after treatment with the GSH and Fer-1. *P < 0.05, ***P < 0.001, n = 3 (biologically independent male mice). B The body weight was recorded for mice in different groups. NS indicates non-significant. C, D The image and weight of the testes from S8/Control, S8/Gss^−/− mice, S8/Gss^−/− mice + GSH, and S8/Gss^−/− mice + Fer-1 mice. *P < 0.05, **P < 0.01; n = 5 (biologically independent animals). E The H&E staining of the testes of mice in different groups. Scar bar = 50 μm. F Immunofluorescence staining of DDX4 for testis. Scar bar = 50 μm. G The number of cells positive for DDX4 per seminiferous tubule. H, I Immunofluorescence staining of γH2AX and the statistical results of positive cells in tubules. G, I ***P < 0.001, n = 3 (biologically independent animals); more than ten tubules were examined for each mouse. J, K The H&E staining image of sperms from cauda epididymis and the percentage of malformed sperms per the H&E staining results. Scar bar = 20 μm. **P < 0.01, ***P < 0.001; n = 4 (biologically independent animals). L Different types of abnormal sperms were counted, including abnormal heads, coiled tails, and decapitated sperms. **P < 0.01, ***P < 0.001; NS indicates non-significant, n = 4 (biologically independent animals). M, N The PNA was used to stain the acrosome of sperms for determining the percentage of sperms with normal acrosome. Scar bar = 20 μm. ***P < 0.001, n = 4 (biologically independent animals); more than 200 sperms were counted for each mouse.