Fig. 3: Aβ-induced APEL is independent of increase in ROS production, decrease in cell viability, apoptotic activation, or endocytosis.
From: Endothelial leakiness elicited by amyloid protein aggregation
a Treatment of HMVECs with Aβo and Aβs (30 min) revealed increases in leakiness with increased concentration. Thrombin (3 U/mL) acted as positive control of induced leakiness. Results are shown as mean ± SD (n = 3 biologically independent samples), analyzed via two-way ANOVA with Tukey’s multiple comparison tests. P values when compared with control are indicated. b No significant increase in ROS production was detected in HMVECs after treatment with either Aβ (20 µM) for as late as 2 h. Hydrogen peroxide (H2O2, 200 µM) acted as positive control. Results are presented as mean ± SD (n = 3 biologically independent samples) and analyzed via one-way ANOVA with Tukey’s multiple comparison tests. P values for comparisons with respective control of same duration are shown. c Induced leakiness under Aβ was not significantly reduced with prior treatment of ROS scavenger NAC (10 mM, 1 h). Results are presented as mean ± SD (n = 3 biologically independent samples) and analyzed via two-tailed Student’s t tests. P values for comparisons with respective non-NAC-treated groups are shown. d No decline in HMVEC viability was detected after treatment with different Aβ proteins (20 µM) for as late as 2 h. Results are presented as mean ± SD (n = 3 biologically independent samples) and analyzed via one-way ANOVA with Tukey’s multiple comparison tests. P values compared with respective control of same time duration are shown. e Immunoblotting revealed no activation of apoptotic markers within concentrations used for Aβ APEL. Staurosporine (STS; 1 µM, 2 h) served as positive control for apoptosis. A representative blot is presented, out of n = 3 biologically independent experiments. f Inhibition of endocytosis in HMVECs (mixture of 5 mM MβCD and 10 µM MDC) did not result in a significant reduction in Aβ APEL. Data are presented as mean ± SD (n = 3 biologically independent samples) and analyzed via two-tailed Student’s t tests. P values compared with respective non-inhibitors-treated groups are shown.
