Figure 3
From: Anti-LRP/LR specific antibodies and shRNAs impede amyloid beta shedding in Alzheimer's disease
Effects of IgG1-iS18.
(a) Aβ levels in HEK293 and SH-SY5Y cells after treatment with IgG1-iS18 and IgG1-HD37 as detected by an Aβ ELISA after 18 hours of antibody incubation. Data shown (mean ± s.e.m) representative of three independent experiments (performed in triplicate) per cell line. *p < 0.05, **p < 0.01, ***p < 0.001, NS not significant; Student's t-test. (b) Aβ concentrations after SH-SY5Y cells were treated with varying doses of IgG1-iS18 for 18 hours, as determined by an Aβ ELISA. Data shown (Mean ± s.d.) comparing Aβ levels of untreated cells (0 μg/ml) and IgG1-iS18 treated cells (25–100 μg/ml), ***p < 0.0001; n = 3; one way ANOVA. (c) Flow cytometric analysis of APP, β-secretase and γ-secretase levels on the surface of HEK293 cells post treatment with IgG1-iS18 (mean ± s.d., NS not significant, n = 3, Student's t-test). (d) (i) Western blot showing sAPPβ levels from cell culture medium after SH-SY5Y cells were treated with varying concentrations (0–100 μg/ml) of IgG1-iS18 for 18 hours. Western blot band intensities from three independent experiments were quantified using Quantity One 4.6 software. Gels have been cropped for clarity and conciseness purposes and have been run under the same experimental conditions. (d) (ii) Obtained band intensities were subsequently used to determine the percentage downregulation of sAPPβ. Data shown (mean ± s.d); ***p < 0.0001; One way ANOVA.
