Additional file 6: of High-density lipoproteins suppress Aβ-induced PBMC adhesion to human endothelial cells in bioengineered vessels and in monoculture Jérôme Robert Emily Button Sophie Stukas Guilaine Boyce Ebrima Gibbs Catherine Cowan Megan Gilmour Wai Cheng Sonja Soo Brian Yuen Arvin Bahrabadi Kevin Kang Iva Kulic Gordon Francis Neil Cashman Cheryl Wellington 10.6084/m9.figshare.c.3860566_D6.v1 https://springernature.figshare.com/articles/journal_contribution/Additional_file_6_of_High-density_lipoproteins_suppress_A_-induced_PBMC_adhesion_to_human_endothelial_cells_in_bioengineered_vessels_and_in_monoculture/5335531 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) 2017-08-22 05:00:00 Alzheimer’s disease Beta-amyloid High-density lipoprotein Engineered vessel Endothelial cells