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Fig. 1 | Epigenetics & Chromatin

Fig. 1

From: Specific functions of TET1 and TET2 in regulating mesenchymal cell lineage determination

Fig. 1

DNA demethylation promotes BMSC differentiation. a Cultured BMSC were treated with 2 μM 5-Azacytidine-2 (5-Aza2) or vehicle control (Veh) under osteogenic inductive conditions for 3 weeks. Mineralised deposits were stained with Alizarin red, and extracellular calcium levels were measured normalised to DNA content per well. b Cultured BMSC were treated with 5-Aza or Veh under adipogenic inductive conditions for 3 weeks. Lipid formation was assessed by Nile red staining, and the number of adipocytes was expressed as a percentage relative to total DAPI+ cells. Data represent n = 3 BMSC donors, *p < 0.05, Students t test. ce BMSC cultured under osteogenic inductive (Osteo) or normal growth (Cont) conditions for 1 to 3 weeks and then analysed by qPCR to assess c TET1, d TET2 and e TET3 gene expression levels, relative to β-actin. fh BMSC cultured under adipogenic inductive (Adipo) or Cont conditions for 1 to 3 weeks and then analysed by qPCR to assess f TET1, g TET2 and h TET3 gene expression levels relative to β-actin. Data represent mean S.E.M, n = 3 BMSC donors, *p < 0.05, one-way ANOVA with multiple comparison analyses. Human BMSC were cultured in Cont media or under Osteo or Adipo inductive conditions for 1 week. Genomic DNA was purified and global levels of i 5hmC and j 5mC were measured using by ELISA relative to total input DNA. Data represent n = 3 BMSC donors, *p < 0.05, Students t test. k Human BMSC were cultured under Cont, Osteo or Adipo inductive conditions for 2 weeks and then analysed by Western blot to assess TET1 and TET2 protein levels compared to Raptor as loading control

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