Moreover, luciferase assay, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay validated that the ligand-bound AR activated Ube2b transcription via direct binding to the androgen-responsive element of the Ube2b promoter. The upregulation of Ube2b gene by testosterone was further demonstrated by Western blot and qRT-PCR in TM4 cells, a mouse Sertoli cell line. The expression of UBE2B was decreased in the testes of Sertoli cell AR knockout (S-AR −/y) mice analyzed by quantitative RT-PCR (qRT-PCR) and immunofluorescence. Here, we identified ubiquitin-conjugating enzyme E2B ( Ube2b) as a critical target gene of AR. Many genes are regulated by androgen and its receptor (AR), but the direct target genes of AR, especially those involved in spermatogenesis and male infertility, remain unclear.
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