FACS-array profiling in retinal endothelial cells from living mouse retinas without pericyte coverage

Pericytes confer vascular stability in the retina, and the loss of pericytes can cause the blood-retina barrier breakdown seen in diabetic retinopathy. To identify endothelial-specific genes expressed in pericyte-deprived retinal vessels, we purified genetically labeled endothelial cells from Tie2-GFP transgenic mice treated with neutralizing antibody against PDGFRb (APB5) and performed gene expression profiling using DNA microarray. To find out endothelial-specific genes associated with the loss of pericyte coverage, the comparison of microarray data was carried out between retinal endothelial cells (data from GSE27238) and APB5-treated retinal endothelial cells. Nine arrays are included. Utilizing fluorescence-activated cell sorting (FACS), we isolated retinal endothelial cells from P8 Tie2-GFP transgenic mice treated with intraperitoneal injections of neutralizing antibody against PDGFRb (APB5) from P0 to P7. RNA extracts from sorted cells were amplified and then hybridized to Affymetrix MGU74v2 series arrays in triplicate.