Fig 2: MACF1 regulates MSCs osteogenic differentiation through TCF4/miR-335(A), Expression of MACF1, TCF4 and mir-335 in MSCs of ROSA26-MACF1f/f and ROSA26-MACF1f/fPrx1-cre mice, as detected by RT-PCR (mean ?± ?SD, *P ?< ?.05 **P ?< ?.01). MSCs were treated with or without TCF4 siRNA (siTCF4) (B), ALP activity of MSCs from ROSA26-MACF1 f/f and ROSA26-MACF1f/fPrx1-cre mice was detected by ALP staining. MSCs were treated with or without TCF4 siRNA (siTCF4) (C), Quantification of relative integrated optical density (IOD) values of ALP staining using Image-Pro Plus 6.0 software (mean ?± ?SD, *P ?< ?.05 **P ?< ?.01, ***P ?< ?.001) (D–E), Expression of BGLAP and DKK1 in MSCs of ROSA26-MACF1f/f and ROSA26-MACF1f/fPrx1-cre mice, as detected by Western blot and RT-PCR (mean ?± ?SD, **P ?< ?.01, ***P ?< ?.001). MSCs were treated with or without TCF4 siRNA (si-TCF4) (F), Expression of MACF1, Tcf4 and miR-335–5p in primary MSCs of MACF1 conditional knockout mice, as detected by RT-PCR (mean ?± ?SD, *P ?< ?.05 **P ?< ?.01).

Fig 3: Correlation between MACF1 and Wnt signaling pathway in SOP(A), Gene expression profile of MSCs from elderly, and elderly osteoporotic patients (GSE35959) (B), Correlation analysis between Macf1 levels and levels of Wnt related gene in MSCs from elderly and elderly osteoporosis patients (C), ALP staining and of MSCs of 4-, 10- and 20-month male C57BL/6 mice (D), Quantification of relative integrated optical density (IOD) values of ALP staining using Image-Pro Plus 6.0 software (mean ?± ?SD, *P ?< ?.05 **P ?< ?.01) (E), Expression of MACF1 in MSCs of 4-,10-and 20-month male C57BL/6 mice, respectively, as detected by RT-PCR (mean ?± ?SD, *P ?< ?.05 **P ?< ?.01) (F) Correlation analysis between Macf1 levels and levels of ß-catenin and TCF4 in MSCs of 4-, 10- and 20-month male C57BL/6 mice (G), Correlation analysis between Macf1 levels and levels of Alp, Runx2, and Bglap in MSCs of 4-,10-and 20-month male C57BL/6 mice.