Figure 5.

Wnt5a/β-Catenin Signaling Is Involved in miR-194-5p-Mediated Osteogenic Differentiation

(A) Effect of miR-194-5p on the luciferase activity of WT Wnt5a 3′ UTR or Mut Wnt5a 3′UTR after treatment with agomiR-194-5p or antagomiR-194-5p in MSC cells. (B) The level of Wnt5a in the serum of healthy volunteers and osteoporosis volunteers was evaluated using quantitative real-time PCR. n = 50, (C and D) The Wnt5a levels were evaluated by quantitative real-time PCR (C) and western blot (B) after antagomiR-194-5p and agomiR-194-5p treatment. (E) The Wnt5a and β-catenin levels were assessed by western blot after indicated treatment. (F–H) Western blot (F) and quantitative real-time PCR were used to evaluate the osteogenic-related markers Runx2 (G) and OCN (H). (I) ALP staining of MSCs after indicated treatment for 3 days. (J) Quantification of ALP staining of MSCs on day 3. (K) Alizarin red staining of MSCs after 21 days following indicated treatment. (L) Quantification of alizarin red staining of MSCs on day 21. The data are expressed as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.