Fig. 4.

Autophagy was required for GLA-induced Sertoli cell and Leydig cell ferroptosis. The protein expression levels of NCOA4 and FTH1 in testes tissues (A). Representative fluorescence confocal images of LC3B and FTH1 colocalization in testes (B). Representative fluorescence confocal images of LC3B and ACSL4 colocalization in testes (C). The protein expression levels of NCOA4 and FTH1 in Leydig cells and Sertoli cells (D). The protein expression levels of NCOA4 and FTH1 in Sertoli cells and Leydig cells following GLA treatment for 24 h combined with 3-MA, CQ, or Baf-A1 (E). The protein expression levels of NCOA4 and FTH1 in Sertoli cells and Leydig cells following GLA treatment for 24 h combined with si-ATG5 (F). Fe²⁺ content (G). MDA content (H). Representative fluorescence confocal images of LC3B and FTH1 colocalization in Sertoli cells and Leydig cells following GLA treatment for 24 h combined with si-ATG5 (I). “*” indicates significant difference compared with the control group (*P < 0.05, **P < 0.01, and ***P < 0.001). “^#” indicates significant difference between the GLA treatment group and combined treatment group (^#P < 0.05, ^##P < 0.01, and ^###P < 0.001).