Figure 6.

EMP2 expression regulates migration and tube formation of HUVEC endothelial cells. (A) Conditioned media was isolated from WT, shRNA control, OE, and shRNA KD#2 (labeled as KD) ARPE-19 cells exposed to 4 h of hypoxic stress (0.5% O2) and used in a transwell cell migration assay. Numbers of migrated cells were imaged and quantified (B). Three independent experiments were conducted, and statistical significance was established using Student’s t-test (unpaired, two-tailed). Bar diagrams depict mean values and standard error of the mean. Scale bar = 50 µm. (C) The morphology of HUVEC cells was analyzed following growth in WT, shRNA control, OE, and shRNA KD ARPE-19 conditioned media. Coverslips were stained with Calcein AM, imaged, and the number of tubes manually counted in a masked fashion. (D) Quantification of the number of tubes imaged above. Experiments were repeated three times, and statistical significance was established using an ANOVA test/Student’s t-test (unpaired, two-tailed). Bar diagrams depict mean values and standard error of the mean. Scale bar = 50 µm.