Figure 3

Adiponectin uptake and modulation in endothelial cells. A) A significant amount of IR800-labeled adiponectin was found attached or within endothelial cells following the transport experiments by confocal imaging of membranes (green?=?70 kDa dextran, red?=?adiponectin). Maximum projections are shown. B) Quantified adiponectin+?area on membranes; Bend cells bound or contained significantly more adiponectin than MS1 cells by ANOVA of image intensity analysis. C) After 6 hours of full-length adiponectin transport across MS1 cells, media from above the monolayer (top) and below the cells and membrane (bottom) was fractionated by gel filtration FPLC. Limited HMW was found to have crossed the cells. 3 samples were pooled for fractionation. D) No change in Cdh5 gene expression was noted in the cells with adiponectin. Bend and EOMA cells had increased Ocln expression as compared to MS1 cells with adiponectin. These cells also increased Tjp1 expression with adiponectin. E) Nos3 was regulated by adiponectin in EOMA cells. No changes were measured in Cav1. (F) Adiponectin receptors 1 and 2, respectively, were not regulated at the mRNA level by adiponectin treatment except for Adipor2 in EOMA cells. Cadh13 expression was not regulated by adiponectin in the cells. Gene comparisons were made by two-tailed student’s t-test with unequal variance. *p < 0.05 and **p < 0.01 with adiponectin. #p < 0.05 compared to MS1 cells.