Retina Investigation of the Regulation of Roundabout4 by Hypoxia-Inducible Factor-1a in Microvascular Endothelial Cells Rui Tian,1 Zaoxia Liu,1 Hui Zhang,1 Xuexun Fang,2 Chenguang Wang,1 Shounan Qi,1 Yan Cheng,1 and Guanfang Su1 1Department of Ophthalmology, Second Hospital of Jilin University, Changchun, Jilin, China 2School of Life Science, Jilin University, Changchun, China Correspondence: Guanfang Su, De- PURPOSE. We determined if hypoxia-inducible factor-1a (HIF-1a) and Roundabout4 (Robo4) partment of Ophthalmology, Second colocalized in fibrovascular membranes (FVM) from patients with proliferative diabetic Hospital of Jilin University, 218 retinopathy (PDR), and investigated the regulation of HIF-1a on Robo4 in microvascular Ziqiang Street, Changchun, Jilin, endothelial cells under normoxic and hypoxic conditions in vitro. 130021, China;
[email protected]. METHODS. Immunofluorescence and confocal laser scanning microscopy were done to analyze Submitted: March 21, 2014 the colocalization of HIF-1a and Robo4 in the FVM. Expression of HIF-1a was knocked down Accepted: February 27, 2015 by small interfering RNA (siRNA) technology to study its effects on Robo4 expression of human retinal endothelial cells (HREC) and human dermal microvascular endothelial cells Citation: Tian R, Liu Z, Zhang H, et al. (HDMEC) under normoxic and/or hypoxic conditions. Full-length human a gene was Investigation of the regulation of HIF-1 Roundabout4 by hypoxia-inducible transfected into HREC and HDMEC using GFP lentivirus vectors to overexpress HIF-1a under factor-1a in microvascular endothelial normoxic conditions. The HIF-1a and Robo4 mRNA and protein expressions were quantified cells. Invest Ophthalmol Vis Sci. by real-time PCR and Western blot.