Sema4D stimulation does not up-regulate the expression of angiopoietin-2, VEGF-A, or HGF. mRNAs extracted from unstimulated (first lane) and Sema4D-stimulated (second lane) HUVECs, were retrotranscribed and amplified for the angiopoietin-2 (Ang-2), VEGF-A, and HGF transcripts (see “Materials and methods”). Plasmid DNA (third lane) was used as positive control. No increase in expression of these genes was observed on Sema4D stimulation. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) amplification was performed as a control for all the extracted RNAs. No differences were observed; one of these controls is shown.