Figure 7.

Metabolic activity of Stx-treated microvascular endothelial cells. bEnd.3 immortalized mouse cerebral cortex microvascular endothelial cells (A), primary human cerebral cortex microvascular endothelial cells (B), primary human neonatal dermal microvascular endothelial cells (C), and CDC.HMEC-1 immortalized human dermal microvascular endothelial cells (D) were incubated with Stx1 (solid lines) or Stx2a (dashed lines) for 42 h. The toxin containing media was removed and fresh media containing 10% alamarBlue was added. Cells were incubated for an additional 3 h and the fluorescent reduction of alamarBlue was measured every 30 min. The 1 h time point is shown except for subconfluent BMECs which depicts the 3 h time point. Graphs depict toxin-treated cells as a percent of untreated control cells. Results are the average of three individual experiments and error bars correspond to standard deviation of the mean. TNF-α upregulates surface ICAM-1 (E). Human brain endothelial cells were incubated with 10 ng/ml TNF-α for 24 h, stained for surface expression of ICAM-1 (CD54) and analyzed by FACS.