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 5. PTPN6 knockdown abrogated the beneficial effects of METTL14 overexpression on SONFH BMSCs. (A, B) qRT-PCR and western blot validated the efficiency of PTPN6 knockdown in METTL14 overexpression cells. (C, D) The CCK-8 and EdU assay evaluated the reversal effect of PTPN6 knockdown on cell proliferation induced by METTL14 overexpression. Scale bar = 100 μm. (E) The ALP activity assay was used to assess the reversal effect of PTPN6 knockdown on the osteogenesis promoted by METTL14 up-regulation. (F) The effects of sh-PTPN6 on the abilities of osteogenic differentiation and adipogenic differentiation of METTL14 overexpression cells were analyzed by Alizarin red staining and oil red staining. Scale bar = 100 μm. (G) Knockdown of PTPN6 counteracted the regulatory effect of METTL14 overexpression on osteogenic and adipogenic related genes. *P < 0.05, **P < 0.01, ***P < 0.001, n.s is no significance.