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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: Ann N Y Acad Sci. 2011 Nov;1237:58–63. doi: 10.1111/j.1749-6632.2011.06213.x

Figure 1.

Figure 1

(A) Effect of Nocturnin on osteoblastogenesis. This figure shows effect of overexpression of (a) Noc on osteoblast gene marker expression as can be observed, there is a significant suppression of osteoblastogeneis, which is evident from the decrease in (b) ATF4 and (c) Osteocalci ; (d) Osterix we also saw a trend towards suppression in (e) CYP51 when Noc is overexpressed, in MC3T3E1 cells compared to GFP cells. (B) The increase in Noc is confirmed by the Flag tag Western blot along with a NOC protein levels in cells, Actin was used as the loading control. (C) (c’)The long term (20 days with change in media every two days) osteoblast differentiation assay on cells that have Noc knocked down show that there is a significant increase in nodule formation compared to control cells. (c’’) Osteoblast differentiation assay for 7 days showed decreased alkaline phosphatase staining in Noc overexpressed cells compared to control cells (n = 3, * indicates P < 0.05). (D) Alizarin red and Alcian blue staining showing the tibial and fibula bones from Noc control +/− and Noc null.