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. Author manuscript; available in PMC: 2019 Nov 1.
Published in final edited form as: J Cell Biochem. 2018 Jun 19;119(10):8204–8219. doi: 10.1002/jcb.26832

Figure 4. Adam17 expression is down-regulated during osteogenic differentiation.

Figure 4.

MC3T3–E1 cells were induced to differentiate with ascorbic acid and β-glycerophosphate for 28 days. Cells were fixed in paraformaldehy for histochemical detection of alkaline phosphatase at the indicated days of differentiation (day 0, 4, 8, 12, 16, 22 and 28) (A). Adam17 and Runx2 mRNA and protein levels as well as ADAM17 and RUNX2 cellular localization were determined by RT-PCR (B, and down graph C), western blot (D, and down graph E) and immunostaining (F), respectively. Expression of osteoblast phenotypic genes alkaline phosphatase (Alpl) as well as cell cycle marker cyclin D (Ccnd1) were additionally examined by RT-PCR and western blot, respectively. The data shown are representative of three experiments with similar outcomes. Runx2 and Adam17 mRNA (C, right graph) and protein (E, left graph) values were normalized to Gapdh and Actin, respectively. All data are presented as mean ± SEM from three independent experiments. *P<0.05 and **P<0.01.