Fig. 4

Osteoprogenitor cell numbers and TET1/2 gene expression are reduced in osteoporotic bones. Ovariectomy (OVX) or sham surgeries (Sham) were performed on 12-week-old C57/BL6 female mice. a Trabeculae in femora harvested 12 weeks post-OVX or Sham treatment analysed by μCT. b The number of clonogenic stromal cells (CFU-F) was assessed from bone marrow and crushed femoral bone isolated from OVX or Sham mice, 12 weeks post-treatment. c Gene expression levels of TET1, TET2 and TET3 normalised to β-actin were measured by qPCR, in stromal cells isolated from the femora of OVX or Sham mice, 12 weeks post-treatment. d Immunohistochemical analysis of 5 μM tibial sections stained with anti-5hmC antibody, in OVX or Sham mice, 12 weeks post-treatment. 5hmC staining was detected on bone surfaces in primary spongiosa and chondrocytes in growth plate region. e Cultured stromal cells derived from the femora of OVX mice and Sham control mice were cultured in normal media, genomic DNA was purified and global levels of 5hmC and 5mC were measured by ELISA relative to total input DNA. Data represent mean S.E.M., n = 5 mice per condition donors, *p ≤ 0.05, two-tailed Student t test