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Firstly, time courses of aggregation were observed in 0 120 min, for reaction consisting of 4 µM SOD1, 2 mM Asc and 7.5 µM λDNA.
In contrast, although the SOD1 can associate with DNA under neutral conditions [47], any observable aggregation phenomenon under the TEM is not found without Asc, as indicated with DSL in examining the time courses of aggregation, revealing that Asc-induced SOD1 oxidation is a key factor in the enhanced aggregation propensity of WT SOD1.
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To assess the changes in MET16 secondary structure over the course of aggregation we used CD spectroscopy in the presence of various cosolutes leading to equal populations of the monomer hairpin conformation φN at t = 0.
We also tested the capability of the assay to capture the full time course of aggregation in the absence of seed fibrils.
DOI: http://dx.doi.org/10.7554/eLife.01681.008 Since our aim is to target the toxic oligomer, we determined when the toxic oligomeric species was present during the course of aggregation.
Panels D F of Figure 1 show the effects of substoichiometric amounts of recombinant lipid-free and lipidated apoE, as well as lipoproteins derived from cultured astrocyte cells, on the time course of aggregation of TMR-Aβ1 42. TMR-Aβ1 42
Besides intra-particle mesopores, over the course of the aggregation process an inter-particle (textural) large pore system is generated.
A change in the electrochemical behavior of silver sols in the time course of particles aggregation and growth was interpreted in agreement with the data of UV vis spectroscopy and electron microscopy observations.
The CD spectra, obtained as described over the time-course of MET16 aggregation, were compiled to a data matrix with the use of the CCA+ software, publically available from http://www.chem.elte.hu/departments/protnmr/cca/.
Note that the very short time-frame that cells were in suspension without cell-cell contact was much less than the time required for detached cells to undergo anoikis [28]; in addition, DAPI staining of cell nuclei during the time course of cell aggregation did not reveal evidence of apoptosis (KK, unpublished results).
In an agreement with previous studies, this peak decreased in the course of AS aggregation.
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