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Fig. 1 | Alzheimer's Research & Therapy

Fig. 1

From: Imipramine and olanzapine block apoE4-catalyzed polymerization of Aβ and show evidence of improving Alzheimer’s disease cognition

Fig. 1

Development of an apoE4-Aβ fibrillization assay for HTS. a Concentrations of Aβ42, apoE4, and ThT were varied in a ½ fraction factorial experiment in a 96-well plate and ThT was measured in relative fluorescence units (r.f.u.) in n = 3 wells per group. b Concentrations of Aβ42, apoE4, and ThT were varied in a response surface experiment in a 384-well plate in n = 3−4 wells per group. c Concentrations of Aβ42, apoE4, and ThT were varied in a second response surface experiment in a 384-well plate in n = 6 wells per group. The experiment was replicated twice, and the results were combined. ac Several representative groups were plotted, and the complete results are provided in Additional file 6. d The quantity of Aβ42 was plotted against the AUC of ThT fluorescence in r.f.u.*hours (h). Linear regression was performed to identify a best-fit line (R2 = 0.53). Two assay conditions that maximized the AUC were identified (red dots). e The Aβ42 to apoE4 ratio was plotted on a log scale against the fold-change in ThT fluorescence. Linear regression was performed to identify a best-fit line (R2 = 0.14). Two assay conditions that maximized the fold-change were identified (red dots). f In the second response surface experiment, the optimal concentrations of Aβ42, apoE4, and ThT that maximize both the AUC (r.f.u.*h) and the fold-change in ThT fluorescence were identified (red lines). g The specificity of the optimized apoE4-Aβ fibrillization assay was evaluated for Aβ42, Aβ40, and a scrambled Aβ42 peptide (Aβscr), with and without apoE4, or for ThT only. The data represent the mean ± SD of n = 3 wells per group. ***P < 0.001 by one-way ANOVA. h The effect of each human apoE isoform, or of apoA-I, at 1 nM concentration on Aβ42 fibrillization was evaluated. The experiment was replicated twice, and the results were combined. The data represent the mean ± SD of n = 7 wells per group. **P < 0.01 by one-way ANOVA. i The effect of apoE isolated from human plasma at different concentrations on Aβ42 fibrillization was compared with that of recombinant apoE4. The data represent the mean ± SD of n = 4 wells per group

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