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. 2019 Jun 12;17:223–234. doi: 10.1016/j.omtn.2019.06.001

Figure 5.

Figure 5

Reduction of PTEN Facilitated Osteoblast Differentiation via Activating PI3K/AKT Signaling

(A) Western blot assessment of PI3K and AKT expression after transfection, using Lipofectamine control, agomiR-NC, agomiR-26a-5p, antagomiR-NC, or antagomiR-26a-5p transfection for 48 h. (B) Western blot assessment of PTEN, PI3K, and AKT levels after transfection with siRNA-NC, siRNA-PTEN or appropriate controls for 48 h. (C) Western blot analysis assessment of p-AKT, AKT, p-PI3K, and p-PI3K levels after transfection using control, 200 μM siRNA-NC, siRNA-PI3K, or siRNA-AKT for 48 h. (D–G) qPCR analysis of Col1a1 (D), ALP (E), OCN (F), and Runx2 (G) expression in MC3T3-E1 cells after transfection with control, siRNA-NC, siRNA-PI3K, or siRNA-AKT for 48 h. (H) Western blot analysis of Col1a1, ALP, OCN, and Runx2 levels after transfection with control, siRNA-NC, siRNA-PI3K, or siRNA-AKT for 48 h. (I) Alizarin red-mediated staining for calcium deposition in MC3T3-E1 cells after transfection with, siRNA-NC, siRNA-PI3K, or siRNA-AKT for 21 days. Scale bar, 10 mm. (J) ALP staining of MC3T3-E1 cells after transfection with control, siRNA-NC, siRNA-PI3K or siRNA-AKT for 48 h. Scale bar, 10 mm. Data are means ± SD of triplicate experiments. *p < 0.05, **p < 0.01, ***p < 0.001.