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. 2011 Nov 22;21(11):1936–1947. doi: 10.1089/scd.2011.0422

FIG. 8.

FIG. 8.

Relation between DLX3 and BMP2 expression during osteogenic differentiation in DFCs. (A) Western Blot analysis with pSMAD1 and β-Actin specific antibodies and protein lysates of DFCs, 72 h after transfection with either pDLX3 or DLX3 siRNA in basal medium. (B) ALP activity (day 10 of differentiation in ODM) in DFCs after DLX3 overexpression and selectively inhibition of the BMP-2 pathway with a specific anti-BMP2 neutralizing antibody. The effect was compared with DLX3- transfected DFCs in ODM and an unspecific antibody (IgG) and with DLX3-transfected DFCs in ODM (Ctr.) without antibody treatment. Results are relative ALP activities to the mean activity of the control culture. All values are means plus standard error (σ/√n) of 4 biological replicates per group. (C, F) The ALP activity was quantified in DFCs after 10 days of BMP2 induction and after transfection with either pDLX3 (C) or DLX3 siRNA(6) (F). All values are means plus standard error (σ/√n) of 3 biological replicates per group; the differences were not significant. Relative gene expression of ALP was determined 3 days after induction with BMP2 and transfection with (D) pDLX3 and pEV or a (G) DLX3 specific siRNA(6). DLX3 expression was determined in DFCs after 3 days osteogenic differentiation, induced with BMP2 or ODM, and transfection with pDLX3 and pEV (E) or DLX3 siRNA(6) and NS siRNA (H). All values are means plus standard error (σ/√n) of 3 biological replicates.