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Figure 6 | Acta Neuropathologica Communications

Figure 6

From: Oligomer-targeting with a conformational antibody fragment promotes toxicity in Aβ-expressing flies

Figure 6

Effect of KW1 on the metabolic activity of Aβ-species on SH-SY5Y cells. (A) MTT assay of SH-SY5Y cells treated with Aβ(1–40) aggregates after 5 days of incubation with or without antibody fragments at 5:1 molar ratio of Aβ versus B10 or KW1. Molar ratio refers to the polypeptide chains. Cells treated with buffer were set to 100%. The general cell toxin staurosporine (STS, 2 μM) served as a positive control. (B,C) MTT-reduction (B) and LDH assay (C) of SH-SY5Y cells treated with Aβ(1–40) incubated for different periods of time with KW1 (blue) or without (white). SSP-treated cells defined 100% LDH activity. (D) MTT assay of SH-SY5Y cells treated with Aβ(1–40), Aβ(1–42) or Aβ(1–42)arc peptides incubated in 50 mM HEPES buffer, pH 7.4, 50 mM NaCl at 37°C for 5 days with KW1 (blue) or without (white). n.s.: non-significant, **: p < 0.01, ***: p < 0.001. Error bars represent standard deviation of n = 6. (E) Corresponding Aβ isoforms with or without KW1 plotted based on original life span data (Table 1) and MTT reduction in vitro (Figure 6). Please note the overlap of several data points. The strong correlation between MTT reduction and neurotoxicity in vivo fits with linear regression (R = 0.98).

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