Figure 4.
Overexpressed MIR99AHG abolished the effects of METTL3 on BMSCs osteogenic differentiation.(a-c) Relative METTL3 protein (a-b) and mRNA (c) expression after transfection was measured via Western blot and qRT-PCR. β-actin was used as internal reference. (d) M6A content in BMSCs after knocking down and overexpressing METTL3. (e-f) Effects of overexpressed METTL3 and MIR99AHG on osteogenic differentiation of BMSCs were evaluated by both ALP staining (e) and Alizarin Red S staining (f). Magnification: × 200. Scale bar = 100 μm. (g) Relative mRNA expression of Osx, Col1A1, OCN and RUNX2 after overexpressed METTL3 and MIR99AHG transfection was quantified with qRT-PCR. β-actin was the internal reference. All experiments have been performed independently in triplicate and data were expressed as mean ± standard deviation (SD). +++p<0.001, vs. NC; &&&p<0.001, vs. siNC; ***p<0.001, vs. NC+Vector; ###p<0.001, vs. METTL3+Vector; ^^p<0.01, ^^^p<0.001, vs. NC+MIR99AHG.