biostudies-literatureCai GPHunan Provincial Science and Technology Departmentthe innovation-driven project of Central South UniversityNational Natural Science Foundation of Chinae13178https://www.ebi.ac.uk/biostudies/studies/S-EPMC8828262biostudies-literatureUnknown55(2)<h4>Objectives</h4>DNA N6-methyladenine (N6-mA) demethylase Alkbh1 participates in regulating osteogenic differentiation of mesenchymal stem cell (MSCs) and vascular calcification. However, the role of Alkbh1 in bone metabolism remains unclear.<h4>Materials and methods</h4>Bone marrow mesenchymal stem cells (BMSCs)-specific Alkbh1 knockout mice were used to investigate the role of Alkbh1 in bone metabolism. Western blot, qRT-PCR, and immunofluorescent staining were used to evaluate the expression of Alkbh1 or optineurin (optn). Micro-CT, histomorphometric analysis, and calcein double-labeling assay were used to evaluate bone phenotypes. Cell staining and qRT-PCR were used to evaluate the osteogenic or adipogenic differentiation of BMSCs. Dot blotting was used to detect the level of N6-mA iCell proliferationAlkbh1-mediated DNA N6-methyladenine modification regulates bone marrow mesenchymal stem cell fate during skeletal aging.PMC88282628190081081770877201800330400082020RC20118200081182170902Wang MJiang TJCai GPLiu YLXiao YLuo LPYuan JfalseAlkbh1-mediated DNA N6-methyladenine modification regulates bone marrow mesenchymal stem cell fate during skeletal aging.<h4>Objectives</h4>DNA N6-methyladenine (N6-mA) demethylase Alkbh1 participates in regulating osteogenic differentiation of mesenchymal stem cell (MSCs) and vascular calcification. However, the role of Alkbh1 in bone metabolism remains unclear.<h4>Materials and methods</h4>Bone marrow mesenchymal stem cells (BMSCs)-specific Alkbh1 knockout mice were used to investigate the role of Alkbh1 in bone metabolism. Western blot, qRT-PCR, and immunofluorescent staining were used to evaluate the expression of Alkbh1 or optineurin (optn). Micro-CT, histomorphometric analysis, and calcein double-labeling assay were used to evaluate bone phenotypes. Cell staining and qRT-PCR were used to evaluate the osteogenic or adipogenic differentiation of BMSCs. Dot blotting was used to detect the level of N6-mA i2022-01-01T00:00:00Z2022 Feb2025-04-25T17:37:21.49Z2025-04-25T17:37:21.49ZS-EPMC88282623501868310.1111/cpr.13178