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. Author manuscript; available in PMC: 2012 Aug 4.
Published in final edited form as: FEBS Lett. 2011 Jun 26;585(15):2410–2418. doi: 10.1016/j.febslet.2011.06.024

Figure 3. APR3 inhibition of osteoblastic proliferation was enhanced by co-overexpression of NELL-1.

Figure 3

A. APR3 and NELL-1 expression plasmids or empty vectors were cotransfected into Saos2 cells. MTS assay was performed at indicated time points.

B. APR3 siRNA increased Saos2 cell proliferation.

C. Effects on mRNA level of Cyclin D1 in Saos2 cells were examined by real-time PCR. APR3 and NELL-1 interaction downregulated Cyclin D1 significantly.

D. Effects on Cyclin D1 protein level were tested by western blot 48 hours after APR3 and NELL-1 transfection in Saos2 cells. Cyclin D1 was dramatically downregulated after APR3 and NELL-1 co-transfection.

E. Parallel experiment in U2OS cells. Cell proliferation was decreased when APR3 and NELL-1 were overexpressed.

F. Endogenous APR3 knockdown increased U2OS cell proliferation.

G. The mRNA level of Cyclin D1 in U2OS cells increased under APR3 silencing condition. Samples were prepared in triplicate. (* p < 0.05, ** p < 0.01, *** p < 0.005)