Please click here to obtain permission to reproduce this image.
Click on image to view larger version.
Figure 3 Impact of Hcy on CD40 Protein Expression in HUVECs
(A) The left panel shows untreated cells. The dashed histogram represents the irrelevant antibody; the green histogram depicts the anti-CD40 antibody. The middle and right panels show HUVECs exposed to respectively 50 µM or 500 µM Hcy for 24 h. Green histograms indicate untreated cells, open histograms depict Hcy-treated cells. Each histogram is representative of at least 3 experiments and shows cell numbers (y axis) versus log fluorescence intensity (x axis) for 6.000 viable cells. (B) Increase in CD40 expression by Hcy in HUVECs is concentration dependent. Cells were exposed to increasing concentrations of Hcy (10, 25, 50, 100, 500 µM) for 12 h, and CD40 expression was evaluated by flow cytometry. Results are from a representative experiment. (C) The graph summarizes results from 3 to 5 different experiments with HUVECs exposed to the indicated concentrations of Hcy for 12 h or 24 h. Data are expressed as mean ± SD of percent of total events. (D) Western blot analysis. The HUVECs were exposed to the indicated concentrations of Hcy for 12 h. The upper panel shows a representative blot. The lower panel depicts mean ± SD of densitometric analyses from 3 separate experiments. Results were normalized for ß-actin. Ctrl = control; FL1 = fluorescent channel 1; Hcy = homocysteine; HUVECs = human umbilical vein endothelial cells.
|
