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. 2021 Mar 13;11(11):5491–5510. doi: 10.7150/thno.55041

Figure 5.

Figure 5

miR-143/145 depletion counteracts the adverse effects of estrogen deficiency on BMSCs function. (A) CFU analysis of colonies in Sham and OVX-induced WT and miR-143/145-/- BMSCs. Lower panel shows the respective quantification. (B) Western blot revealing the protein expression of Nanog, Sox2, Oct4, and Satb2. (C) γH2AX staining showing the senescence difference in WT and miR-143/145-/- BMSCs following OVX. Lower panel shows the respective quantification. Scale bars: 4 μm. (D) Western blot revealing the protein expression of Trp53 and Cdkn1a. (E) Alizarin red revealing the alternation of osteogenic differentiation. Right panel shows the respective quantification. (F) Western blot revealing osteogenic markers of Runx2 and Bglap2 and (G) core TFs and SATB2. (H) The expression of pri-miR-143/145 was increased in OVX-induced BMSCs and in estrogen or siERβ treated BMSCs. (I) ChIP analysis of ERβ binding to the shared putative region of pri-miR-143 and pri-miR-145 promoter. (J) WT and Mutation vectors were constructed for dual-luciferase reporter assay and co-transfected with estrogen or siERβ. Results are presented as the mean ± S.D. *p < 0.05; **p < 0.01; #p > 0.05 by Student's t test and one-way ANOVA.