PAUP (450bp)-Aug12-2009 without Host

Nexus file for generating NJ tree in PAUP

Source modifier table for GenBank-corrected

Source data for sequences and origin of genomic samples for GenBank accession numbers JN800360-JN800398

Bar graph showing the additive effect of β-Amyloid (Aβ) and AChE C-terminus peptides of 30 amino acids (T30) on cell viability.

<p>Cells were treated for 1 hour. Data is presented as % cell viability ± SEM (N = 3). * vs Control: **<i>P</i><0.01, **<i>P<</i>0.001; ^# vs T30: ^##<i>P</i><0.01, ^###<i>P</i><0.001;^& vs Aβ: ^&<i>P</i><0.05, ^&&<i>P</i><0.01.</p

Alignment of the sequences of β-Amyloid (Aβ) compared with the AChE C-terminus peptides of 30 amino acids (T30), 14 amino acids (T14), and the residual 15 amino acid sequence between the two (T15).

<p>Note homology between the 9 amino acid sequence (9AA) possible in human plasma and T14, T30 and Aβ (shaded panels), but not T15.</p

Dose-response curves representing cell viability (% of control) after 1 and 6 h of treatment with AChE C-terminus peptides of 30 amino acids (T30) with concentrations.

<p>Data are presented as the percentage (of control) of the mean ± S.E.M. from five independent experiments carried out on triplicates (*<i>P<</i>0.05, N = 5).</p

Bar graph shows the increase of Acetylcholinesterase (AChE) activity.

<p>Cells were treated for 15 and 60 minutes with Hydrogen peroxide (H₂O₂) 100 µM, β-Amyloid (Aβ) 0.7 µM, AChE C-terminus peptides of 30 amino acids (T30) 0.7 µM and the residual 15 amino acid sequence (T15) 0.7 µM. Data are presented as the percentage of the mean ± S.E.M. divided by the number of extant cells and obtained from independent experiments carried out on triplicates. *<i>P</i><0.05 and ***<i>P</i><0.001 vs Control; # <i>P</i><0.05 between T30 treatments. Dotted line shows basal AChE activity (Control 100%).</p

Effect of AChE C-terminus peptides of 30 amino acids (T30) on morphology of PC12 cells.

<p>(A) Control cells presented a round shape with a big nucleus. In contrast, (B) Cells treated with T30 presented granulation of the nucleus () as apoptotic feature.</p

Representative table of cell viability (% of Control) after 15 and 60 minutes treatments with Hydrogen peroxide (H₂O₂) 100 µM, β-Amyloid (Aβ) 0.7 µM, AChE C-terminus peptides of 30 amino acids (T30) 0.7 αM and the residual 15 amino acid sequence (T15) 0.7 αM. Data are presented as the percentage of the mean ± S.E.M. from five independent experiments carried out on triplicates (N = 5).

*<p><i>P</i><0.05 and **<i>P</i><0.01 vs Control.</p

Bar graph showing lactate dehydrogenase release.

<p>Cells were treated for 1 hour with AChE C-terminus peptides of 30 amino acids (T30) 0.7 µM, the residual 15 amino acid sequence (T15) 0.7 µM and hydrogen peroxide (H₂O₂) 100 µM. Data are presented as the percentage (of control) ± S.E.M. (N = 3). * vs Control: **<i>P</i><0.01.</p

Possible scheme of the effects of T30 on PC12 cells.

<p>The population of cells (1) is treated with AChE C-terminus peptide of 30 amino acids (T30), causing a reduction of 30% of the population (2) and a consequent increase in AChE release from the 70% of the extant population as a ‘compensatory’ effect (3). The released AChE could be cleaved to yield T30 which then would act at the α7-nAChR, completing a feed-forward cycle of toxicity.</p