Fig. 4.

Culture media modifies IL-1β induced inflammatory mediator expression in human brain pericytes. Human brain pericytes were plated at a low density and allowed to proliferate in either DMEM/F12 with 10% FBS or Pericyte Medium for 7 days. For the final 24 h 10 ng/mL IL-1β or vehicle (0.0001% BSA in PBS) was added. Pericytes were fixed and immunostained for the inflammatory mediators ICAM-1 or MCP-1 (a). Nuclei were counterstained with Hoechst. The integrated intensity of ICAM-1/cell was determined by automated image analysis (b), the secretion of soluble ICAM-1 (sICAM-1) was determined by a cytometric bead array (c) and the gene expression of ICAM1 was determined by qRT-PCR (d). Similarly, the integrated intensity of MCP-1/cell was determined by automated image analysis (e), the secretion of MCP-1 was determined by a cytometric bead array (f) and the gene expression of MCP1 determined by qRT-PCR (g). Data are displayed as mean ± SEM from three independent experiments except CBA data which depicts mean ± SEM of triplicate wells from one representative case of three independent experiments. NS = p > 0.05; **p < 0.01; ***p < 0.001 as designated, or compared to vehicle control in respective media (Two-way ANOVA). Scale bar = 50 µm