M-CSF induces VEGF and regulates angiogenesis <i>in vivo</i>.

<p>A) Matrigel™ was resuspended with PBS <i>(PBS)</i>, rmVEGF (10 ng/ml) <i>(VEGF)</i> or rmM-CSF (100 ng/ml) <i>(M-CSF)</i>. The slides were stained with either an anti-mouse VEGF antibody or its isotype IgG. Brown staining represents VEGF within the plug. Pictures were taken using a 20× objective lens. B) The amount of brown stain (VEGF) within the plugs was determined by taking 15 digital images randomly using a 20× objective lens in a blinded manner. Quantification of VEGF within each plug was determined by Adobe Photoshop histogram pixel analysis of the brown stain. C) Matrigel™ was resuspended with PBS <i>(PBS)</i>, rmVEGF (10 ng/ml) <i>(VEGF)</i>, rmM-CSF (100 ng/ml) <i>(M-CSF)</i>, or rmM-CSF (100 ng/ml)+anti-VEGF neutralizing antibody (5 µg/ml) <i>(M-CSF+anti-VEGF neutralizing Ab)</i>. The plugs were stained for endothelial cells using an anti-mouse von Willebrand factor (vWf) antibody. Brown staining represents vWf(+) cells within the plugs. Pictures were taken using a 20× objective lens. D) Quantification of blood vessels within each plug was determined by counting the total number of vWf(+)-stained endothelial cells that formed a complete circle (red arrows) throughout each entire plug using a 20× objective lens in a blinded manner. This data represents three plugs per group.