Fig. 5: Ferroptosis were observed in the testes of 8-month-old S8/Gss−/− mice. | Cell Death & Disease

Fig. 5: Ferroptosis were observed in the testes of 8-month-old S8/Gss−/− mice.

From: Gss deficiency causes age-related fertility impairment via ROS-triggered ferroptosis in the testes of mice

Fig. 5

A The content of GSH in testis from S8/Control and S8/Gss−/− mice at 2 or 8 months. B, C The relative levels of ROS in testis and sperms from 2- or 8-month-old S8/Control and S8/Gss−/− mice. A–C ***P < 0.001; NS indicates non-significant, n = 6 (biologically independent animals). D, E The expression levels of autophagy-related proteins, including LC3, ATG5, ATG7, and P62, in the testis of 2- or 8-month-old mice. *P < 0.05, **P < 0.01, ***P < 0.001; n = 6 (biologically independent animals). F, G Western blotting to determine the apoptosis-related proteins levels, including BCL-2, BAX, cleaved-Caspase 3, and Caspase 3. *P < 0.05, **P < 0.01, ***P < 0.001; NS indicates non-significant, n ≥ 5 (biologically independent animals). H The mRNA levels of anti-oxidant genes (Gpx4, Gsr, Glrx, Nqo1) and pro-oxidation genes (Alox15, Lpo) were examined in the testes of 8-month-old S8/Control and S8/Gss−/− mice. *P < 0.05, **P < 0.01, ***P < 0.001; n = 3 (biologically independent animals). I The protein levels of GPX4 and ALOX15 in the testis were examined in 2-month-old mice. ***P < 0.001; n = 6 (biologically independent animals). J The GPX4 and ALOX15 expressions were determined in testis from 8-month-old S8/Control and S8/Gss−/− mice by western blot. ***P < 0.001; n = 6 (biologically independent animals). K Western blot was used to determine the levels of 3-NT and 4-HNE in the testes of 2-month-old mice. NS indicates non-significant; n = 4 (biologically independent animals). L The levels of 3-NT and 4-HNE in the testes were increased in 8-month-old S8/Gss−/− mice compared to those in S8/Control mice. **P < 0.01, n = 4 (biologically independent animals). M, O The relative levels of 8-OHdG and Fe2+ exhibited no differences between 2-month-old S8/Control and S8/Gss−/− mice. NS means non-significant, n = 3 (J) and 4 (L) biologically independent animals. N, P The relative levels of 8-OHdG and Fe2+ of the testes of 8-month-old mice were determined by the corresponding assay kit. **P < 0.01; n = 3 (N) and 4 (P) (biologically independent animals).

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