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Loss of Atg5 in Sertoli cells enhances the susceptibility of cadmium-impaired testicular spermatogenesis in mice 1 Funding 1 1 Introduction 1 2 Materials and methods 2 2.1 Reagents 2 2.2 Animal treatments 2 2.3 Cell culture 2 2.4 Epididymal sperm counts and sperm motility assays 2 2.5 Histological examination 2 2.6 Extraction of primary Sertoli cells 2 2.7 Western blotting 3 2.8 MTT assay 3 2.9 Immunohistochemistry 4 2.10 Enzyme linked immunosorbent assay (ELISA) 5 2.11 Cell plasmid Transfection 5 2.12 Statistical analysis 5 3 Results 6 3.1 Sertoli cell-specific knockout of Atg5 aggravates cadmium-caused sperm quality decline in mice 6 3.2 Sertoli cell-specific knockout of Atg5 aggravates cadmium-impaired testicular germ cell development 6 3.3 Sertoli cell-specific knockout of Atg5 aggravates cadmium-inhibited testicular meiosis 6 3.4 Sertoli cell-specific knockout of Atg5 aggravates cadmium-disrupted testicular retinoic acid signaling 6 3.5 Sertoli cell-specific knockout of Atg5 aggravates cadmium-reduced the level of WT1 in mice 7 3.6 Overexpression of Wt1 alleviates Cd-inhibited retinoic acid signaling in Sertoli cell 7 4 Discussion 7 CRediT authorship contribution statement 9 D

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EN

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nonfiction