Combination of VEGF and BMP-6 enhances expression of osteogenic genes.

<p>Expression of ALP (A), COL1A1 (B), runx2 (C) and osterix (D) genes were determined by real time PCR at day 7. * denotes p<0.05.</p

The D1 cells are multipotent.

<p>The D1 cells were stained with DAPI and phalloidin to visualize cell morphology (A). Osteogenesis was assayed by staining with Alizarin Red (B) and adipogenesis was assayed by staining with Sudan IV (C).</p

List of antibodies used for fluorescence activated cell sorting.

<p>List of antibodies used for fluorescence activated cell sorting.</p

Enhanced expression of CADM1 in implants of D1 cells preconditioned with VEGF and BMP-6.

<p>RNA was isolated from the harvested implants and converted into cDNA. Using this cDNA as a template, mRNA expression of CADM1 was quantitatively determined in real time PCR. * denotes p<0.05.</p

List of primers used in PCR.

<p>List of primers used in PCR.</p

BMPs do not enhance Smad 1/5/8 phosphorylation and expression of runx2 and osterix genes.

<p>mRNA levels of runx2 (A) and osterix (B) were quantified using real time PCR. Smad phosphorylation was determined by western blots (C) and band intensity was quantified using ImageJ software (D).</p

Preconditioning of BMP-non-responsive D1 cells with VEGF and BMP-6 enhances their ability to induce ectopic bone formation.

<p>The bone density of harvested implants was measured by radiography and Image J software (A-C). Representative images are shown in the figure. H and E staining revealed typical histology of bone (E). Real time PCR showed modulation of RANKL/OPG ratio (D). * denotes p<0.05 compared with OM group at weeks 2.</p

Expression of ALP and COL1 genes and genes for transcription factors osterix and Dlx5 in the implants of D1 cells.

<p>mRNA expression was quantified using gene specific primers and real time PCR. cDNA prepared from RNA isolated from the harvested implants was used as the template. * denotes p<0.05 compared with OM group of the respective time point, unless indicated otherwise by the horizontal lines.</p

List of primers used in real time PCR.

<p>List of primers used in real time PCR.</p