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DNA fragmentation in apoptotic cells was detected using a TUNEL assay kit. Briefly, testis sections were embedded in paraffin, cut into 5-µm-thick sections, and incubated with the TUNEL kit. Cells were fixed in 4% paraformaldehyde solution for 20 min at room temperature. The cells were counterstained with DAPI to mark the nuclei and were then visualized by fluorescence microscopy. Apoptotic cell nuclei appeared green, and overlapping green fluorescence and blue nuclear fluorescence indicated apoptosis.
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