Fig 3. Silencing Ephrin-B2 expression in pericytes decreased diabetes-mediated angiogenic signaling.

Ephrin-B2 was silenced in human brain microvascular pericytes using siRNA technology. Human brain microvascular pericytes and endothelial cells were grown in ratio one to four respectively in normal glucose or high glucose (25 mM) and palmitate (200 uM) to mimic diabetic conditions. L-glucose was used as an osmotic control. Representative images and quantifications from matrigel tube formation and filopodia extensions in pericytes/endothelial co-cultures are shown in panels A and B. Our results shows a significant increase in tube formation and number of filopodia extensions in diabetic conditions compared to control. Silencing Ephrin-B2 expression in pericytes abolished diabetes induced angiogenic signaling in pericytes/endothelial cell co-culture. (Two way ANOVA, Gene vs diabetic condition, significance: yes, Interaction: yes, n = 3 duplicate).