Fig. 6: TET proteins associate with RUNX2 and promote osteogenesis with RUNX2 synergistically. | Nature Communications

Fig. 6: TET proteins associate with RUNX2 and promote osteogenesis with RUNX2 synergistically.

From: TET enzymes regulate skeletal development through increasing chromatin accessibility of RUNX2 target genes

Fig. 6: TET proteins associate with RUNX2 and promote osteogenesis with RUNX2 synergistically.

a Co-immunoprecipitation (Co-IP) of Flag-TET1 or Flag-TET2 or Flag-TET3 with HA-RUNX2. HA-RUNX2 expressing plasmid was co-transfected with Flag-TET1 or Flag-TET2 or Flag-TET3 in 293 T cells. Whole cell lysate was used for immunoprecipitation and then blotted with indicated antibodies. Representative images for two independent samples. b Co-immunoprecipitation (Co-IP) of Flag-TET1-CD or Flag-TET2-CD or Flag-TET3-CD with HA-RUNX2. HA-RUNX2 expressing plasmid was co-transfected with Flag-TET1-CD or Flag-TET2-CD or Flag-TET3-CD in 293 T cells. Whole cell lysate was used for immune precipitation and then immunoblotting with indicated antibodies. Representative images for 3 independent samples. c ALP staining and Alizarin red S staining after osteoblast differentiation for 7 days (left) and 21 days (right), respectively. Cells were infected with Runx2-lentivirus and Tet2-adenovirus separated or combined. Scale bar = 1 mm. d ALP activity quantification was measured by phosphatase substrate assay as A405/Ala.Blue. *P < 0.05, **P < 0.01. Ordinary one-way ANOVA. Data are presented as mean ± s.d., n = 3 independent cell supernatants. e–i RT-qPCR analysis of Runx2 (e), Tet2 (f), Col1α1 (g), Alpl (h), and Bglap (i) expression after osteoblast differentiation for 7 days, cells were from WT and TCKO mice. *P < 0.05, **P < 0.01. Ordinary one-way ANOVA. Data are presented as mean ± s.d., n = 4 independent samples.

Back to article page