Research Paper Volume 11, Issue 17 pp 6983—6998

Figure 6. Knockdown of endogenous LRP6 resembles the effect of miR-126a-3p on osteogenesis via inhibition of the WNT pathway. (A) The mRNA and protein levels of LPR6 were detected by qRT-PCR and western blot assays respectively in LPR6 siRNAs or negative control siRNA-transfected cells. (B) The mRNA levels of osteogenic-related genes were detected by qRT-PCR assay on day 6 of osteogenic differentiation. (C) The protein levels of osteogenic-related genes were analyzed using western blot assays on day 6 of osteogenic differentiation. (D, E) ALP staining and ALP activity analyses were used to indicate the early differentiation on day 6 of osteogenic differentiation. (F) Alizarin red staining was performed to indicate calcium salt deposits on day 12. (G, H) qRT-PCR and Western blot assays analyzed the expression levels of CTNNB1. Scale bars: 200 μm. Quantitative data are presented as the mean ± S.D. (n =3). *P<0.05; **P<0.01; ***P<0.001.