Sodium citrate buffer (0.01 M and pH 6) was used on testis sections with an antigen retrieval step. UltraVision detection system (Thermo-Scientific, USA) was applied in immunohistochemistry assays, as previously described by Urriola-Muñoz, P. et al.22 (link). Slides were incubated overnight at 4 °C with an anti-caspase 3 antibody (Cell Signaling, USA) at 1 mg/ml and the sections counter-stained with hematoxylin and subsequently evaluated under a microscope. Active caspase-3 cells were quantified in a minimum of 100 seminiferous tubules per each replicate, per (n).
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