The mean fluorescence values obtained from the flow cytometry histogram were plotted against incubated Aβ_1–42 concentration in the presence of serum (A) and under serum deprived condition (B).

<p>The mean fluorescence values obtained from the flow cytometry histogram were plotted against incubated Aβ_1–42 concentration in the presence of serum (A) and under serum deprived condition (B).</p

Affects of exogenous Aβ_1–42 on the level of eNOS and autophagy in endothelial cells.

<p>Endothelial cells were treated with 5 µM and 10 µM Aβ_1–42 and incubated for 12 hrs. Protein levels of eNOS, nNOS, caveolin-1 and the autophagy marker LC3B were analyzed by Western blotting (A). (B), quantitation of protein levels of eNOS, nNOS and caveolin-1 normalized to β-actin control. Results are mean ± SD (n = 3–5). (C), quantitation of LC3B II/I ratio normalized to β-actin control. Results are mean ± SD (n = 3). **, P<0.01 compared to control.</p

Aβ_1–40 expression monitored by immunofluorescence microscopy.

<p>Endothelial cells were incubated with different concentrations of fibrils for 12 hrs and processed as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058194#s2" target="_blank">materials and method</a>. Phase contrast (left panels) and fluorescence images (right panel) were taken showing the same field.</p

The morphological changes of Hep-1 cells incubated with Aβ_1–42 in the presence and absence of serum analyzed by scattering data.

<p>The effect of Aβ_1–42 concentration (A, D: 0 µM; B, E: 5 µM; C, F: 10 µM) in both forward and side scatter values is given in the dot plot.</p

Detection of fibrillar Aβ_1–42 induced Aβ_1–40 production by flow cytometry.

<p>For the analysis of Aβ_1–40 synthesis in amyloidic condition, Hep-1 cells were treated with fibrillar Aβ_1–42 in the presence (A) or absence (B) of serum, stained with anti Aβ_1–40 antibodies (biotinylated) and streptavidin conjugated phycoerythrin. Histograms are shown for the control with no added amyloid (ctrl) and for cells incubated with various concentrations of Aβ_1–42.as indicated in the figure.</p

Fibrillar Aβ_1–42 induced APP upregulation in Hep-1 cells.

<p>Morphology of fibrillar Aβ_1–42 was confirmed by TEM (A).Cells were untreated or treated with 2 µM, 10 µM, 20 µM Aβ_1–42 for 24 hrs. Cell lysates were subjected to Western blot analysis using 8E5 antibody (B). The blot was quantitated by densitometric analysis (C). The fold increase of APP production compared to untreated control cells were shown as mean ± SD (n = 4). *, P<0.05; **, P<0.01 compared to control.</p