Figure 3: Dicer disruption does not affect dexamethasone inhibited proliferation and differentiation in vitro.

(A) Primary osteoblasts were isolated from the calvaria of DicergtRosa26CreERT2 mice. Deletion of Dicer was induced by treating the cells with 1 μM 4-hydroxytamoxifen (4-OHT) for 3 consecutive days. After deletion, cells were treated with or without 10−6 M Dexamethasone (Dex) for 6 days. Cell numbers were assessed by automated microscope. (B) Osteoblast differentiation and mineralization of calvaria-derived primary osteoblasts of DicergtRosa26CreERT2 mice was assessed by alkaline phosphatase (day 10) and alizarin red (day 20) staining after Dicer deletion with 4-OHT and differentiation for 10 and 20 days, respectively. (C) Quantification of ALP activity was done by automated microscope. (D,E) Quantitative mRNA expression analysis of Col1a1 (D) and Bglap (E) measured by qRT-PCR in calvaria-derived primary cells differentiated for 10 days or 20 days, respectively (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, n.s. not significant, OI vs OI + Dex.