biostudies-literatureWeil PPWitten/Herdecke University ‘Interne Forschungsförderung’30564https://www.ebi.ac.uk/biostudies/studies/S-EPMC11659419biostudies-literatureUnknown14(1)Testicular cell differentiation is a highly regulated process, essential for male reproductive health. The histone variant H3.5 is apparently a critical player in this intricate orchestra of cell types, but its regulation and function remains poorly understood. To elucidate its role, we fractionized testicular cells using c-Kit/CD117 as a separation marker and analyzed H3.5 expression. Further, we investigated the regulation of H3.5 expression using public data repositories. We explored DNA methylation patterns in specific regions of the H3-5 gene and assessed H3-5 copy number gain in seminoma specimens. Additionally, we examined the testicular localization of H3.5 and its histone chaperone interactions to understand its regulation at the protein level. We used qRT-PCR, MeDIP, and qPCR to Scientific reportsHistone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones.PMC11659419IFF2018-56 AND IFF 2023-69Wirth BDegener SOetjen EGodde DCremers JFCzarnetzki MPostberg JPembaur ABraun SZirngibl KWeil PPBusscher HfalseHistone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones.Testicular cell differentiation is a highly regulated process, essential for male reproductive health. The histone variant H3.5 is apparently a critical player in this intricate orchestra of cell types, but its regulation and function remains poorly understood. To elucidate its role, we fractionized testicular cells using c-Kit/CD117 as a separation marker and analyzed H3.5 expression. Further, we investigated the regulation of H3.5 expression using public data repositories. We explored DNA methylation patterns in specific regions of the H3-5 gene and assessed H3-5 copy number gain in seminoma specimens. Additionally, we examined the testicular localization of H3.5 and its histone chaperone interactions to understand its regulation at the protein level. We used qRT-PCR, MeDIP, and qPCR to 2024-01-01T00:00:00Z2024 Dec2025-04-04T01:09:22.204Z2025-04-04T01:09:22.204ZS-EPMC116594193970277710.1038/s41598-024-83206-9