VEGFB treated macrophage

VEGFB effect in resting and active macrophage. The goal of this study was to investigate the effects of Vascular Endothelial Growth Factor B (VEGFB) on gene expression in resting and active macrophages. Resting macrophages were obtained from peripheral blood of healthy volunteers. A part of macropahges were activated by Lipopolysaccharides (LPS). Both types of cells were treated by VEGFB. Total RNA was extracted using conventional Trizol extraction, quantified by Nanodrop and analyzed by Bioanalyzer. 1microg of RNA was amplified using AminoAllyl MessageAmp kit. 5µg amino allyl-coupled RNA was labeled with Cy3 or Cy5 dyes. Dye coupling yield >5% was a prerequisite for further analysis. 750ng of labeled RNA was hybridized on 25K gene microarrays for 17 hours at 60°C. 4 arrays per sample were hybridized and scanned with the Genepix 4000B Scanner (Molecular Devices). Microarray data quantification and pre-processing was performed with the MAIA software . The SAM tool was applied to identify differentially expressed genes. Macrophages from 2 different volunteers were treated by vehicle, or VEGFB, or LPS, or LPS and VEGFB