Figure 3: The E22P-Aβ42 dimer forms stable 6~8-mer associated with neurotoxicity. | Scientific Reports

Figure 3: The E22P-Aβ42 dimer forms stable 6~8-mer associated with neurotoxicity.

From: Monoclonal antibody with conformational specificity for a toxic conformer of amyloid β42 and its application toward the Alzheimer’s disease diagnosis

Figure 3

(a) Th-T assay result for each Aβ (25 μM) after incubation for the indicated duration at 37 °C. ●, the E22P-Aβ42 dimer; ○, the biotinylated E22P-Aβ42 dimer; ▲, E22P-Aβ42; △, biotinylated E22P-Aβ42. Data are expressed as mean ± s.e.m. (n = 8). (b) TEM analysis of Aβ aggregates formed from E22P-Aβ42 and E22P-Aβ42 dimer after incubation for the indicated duration at 37 °C. Arrowheads indicate oligomers. Scale bar = 50 nm. Upper, E22P-Aβ42; lower, the E22P-Aβ42 dimer. (c) Analysis of secondary structure of E22P-Aβ42 and the E22P-Aβ42 dimer (25 μM) after incubation for the indicated duration at 37 °C using CD spectrometry. (d) The ability of E22P-Aβ42 and E22P-Aβ42 dimer (25 μM) to form soluble oligomers was evaluated by size exclusion chromatography after incubation for the indicated duration at 37 °C. The peptide was detected by absorbance at 220 nm. Molecular marker sizes are shown in kDa. V0: void volume. (e) The viability of SH-SY5Y cells treated with either E22P-Aβ42 or E22P-Aβ42 dimer at the indicated concentration at 37 °C for 16 h. Data are expressed as the mean ± s.e.m. (n = 3). Absorbance obtained after adding vehicle (Veh: 0.1% NH4OH) was taken as 100%. #p < 0.05 vs vehicle alone, *p < 0.05.

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