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. 2018 Dec 4;9(6):1103–1121. doi: 10.14336/AD.2018.1110

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Copyright: © 2018 Junfen et al.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 8. — miR-1292 regulates hADSC senescence and osteogenic differentiation through Wnt/β-catenin signaling. (A) The Wnt/β-catenin inhibitor XAV939 was used to block this pathway. The expression of senescence-associated markers and osteogenesis-related genes was analyzed by qRT-PCR after different treatments. (B) Western blot was performed to analyze the protein levels of senescence markers, osteogenic factors, and β-catenin after treatment with XAV939. (C) ALP staining and activity assays were used to analyze the effects of different concentrations of XAV939 on osteogenesis. (D and E) Western blot was performed to analyze the protein levels of FZD4 and β-catenin after miR-1292 overexpression or FZD4 downregulation in hADSCs. (F and G) ALP staining and ALP activity were measured on day 4 of osteogenic differentiation. (H) Alizarin red staining was performed to indicate calcification. (I) The protein levels of senescence-associated and osteogenic factors were detected by western blot. (J) qRT-PCR was performed to analyze the mRNA levels of senescence- and osteogenesis-related genes after different treatments. GAPDH was used as an internal control for western blot. qRT-PCR data were normalized to PPIA or GAPDH expression; results are presented as the mean ± SD, n = 3. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 compared to the control. Scale bars: 200 μm.