Additional file 6: of High-density lipoproteins suppress Aβ-induced PBMC adhesion to human endothelial cells in bioengineered vessels and in monoculture

Adhesion molecules are enhanced by TNF-α but not Aβ.(a-c) hCMEC/D3 were primed with 100 μg/mL HDL for 2 h followed by stimulation with 1 ng/mL TNF-α for 3 h. Cell lysates were prepared in RIPA and protein levels of (a) ICAM-1 and (b) VCAM-1 were measured by denaturing immunoblotting (c). (d-g) hCMEC/D3 were stimulated with monomeric (d,f) Aβ40 or (e,g) Aβ42 at the indicated concentrations and (d,e) ICAM-1 and (f,g) VCAM-1 mRNA levels were measured by real-time PCR. (h-j) Following Aβ stimulation, cell surface proteins were biotinylated and isolated by immunoprecipitation. Protein levels of cell surface and total (h) ICAM-1 and (i) VCAM-1 were measured by denaturing immunoblotting (j). (k-m) hCMEC/D3 were pre-treated with 100 μg/mL of HDL for 2 h followed by stimulation with Aβ. After 3 h total and cell surface ICAM-1 expression were measured as above. Graphs represent means ± SD from at least 3 independent trials. ***p < 0.001 versus vehicle, § p < 0.05, versus Aβ or TNF-α (PDF 2282 kb)