Figure 5 | Scientific Reports

Figure 5

From: Endocytic uptake of monomeric amyloid-β peptides is clathrin- and dynamin-independent and results in selective accumulation of Aβ(1–42) compared to Aβ(1–40)

Figure 5

Uptake of Aβ(1–40), Aβ(1–42) and Trf in SH-SY5Y cells under conditions that perturb actin dependent endocytosis, macropinocytosis and Arf6-dependent endocytosis. (a) Uptake of HF488-labelled Aβ(1–40), Aβ(1–42) and AF647-labelled Trf in cells treated with 10 µM cytochalasin A (cyto A) and 25 µg/ml cytochalasin D (cyto D) to perturb actin polymerisation, or 10 µM IPA-3 and 25 nM wortmannin (wort) to perturb macropinocytosis. The peptide uptake is reported as mean cellular uptake relative to control (cells not treated with inhibitor) calculated based on mean cellular fluorescence intensity ± SD of the total number of gated live cells for three replicate samples (n = 3) measured by flow cytometry and corrected for baseline contributions by subtracting the cellular autofluorescence. (b,c) Images showing uptake of HF488-labelled Aβ(1–40) (b) and Aβ(1–42) (c) (green) in cells transfected with BFP-labelled Arf6 WT or Q67L (blue) alongside transmitted images showing the cells. The cells were incubated with Aβ 24 h post transfection. The Aβ concentration was 1 μM and the concentration of Trf was 5 µg/ml in all experiments. Cells were incubated with Aβ for 1 h (flow cytometry) or 24 h (confocal microscopy), and Trf for 5 min.

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