Research Paper Volume 13, Issue 24 pp 25903—25919

METTL14 benefits the mesenchymal stem cells in patients with steroid-associated osteonecrosis of the femoral head by regulating the m6A level of PTPN6

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Figure 6. PTPN6-mediated METTL14 activated the Wnt signaling pathway. (A) The activity of Wnt signaling in SONFH BMSCs was analyzed by TOP/FOP-Flash reporter assay. (B) The protein level of β-catenin in the nucleus was measured by western blot. (C) Western blot detected the total and phosphorylated β-catenin levels in SONFH BMSCs treated as indicated. (D) Immunofluorescence staining was used to analyze the distribution of β-catenin in the cytoplasm and nucleus after METTL14 overexpression and PTPN6 knockdown. Scale bar = 50 μm. (E) The interaction of PTPN6 and GSK-3β in BMSCs was verified by co-IP assay. (F) The effect of PTPN6 inhibition on the phosphorylation level of GSK-3β in SONFH BMSCs was determined by western blot. *P < 0.05, **P < 0.01, ***P < 0.001, n.s is no significance.