Inhibition of proteasomal degradation lowers αsyn solubility.

Abstract

<p>(<b>A</b>) Quantitative analysis of GFP fluorescence of cells expressing αsyn-GFP<sub>11</sub>+ GFP<sub>1–10</sub> (blue) and TP αsyn-GFP<sub>11</sub>+ GFP<sub>1–10</sub> (red). Relative fluorescence was calculated by normalizing the fluorescence of cells 12 and 24 hrs post transfection to the fluorescence measured at 0 hr. *<i>p</i><0.005; **<i>p</i><0.05. (<b>B</b>) Relative fluorescence of cells expressing αsyn-GFP<sub>11</sub> and GFP<sub>1–10</sub> (blue) and TP αsyn-GFP<sub>11</sub>+ GFP<sub>1–10</sub> (red), 24 hrs post transfection. Cells were incubated for 24 hrs with increasing concentrations of lactacystin (0–5 µM). Relative fluorescence was evaluated by normalizing the fluorescence of treated cells to the fluorescence of untreated cells. *<i>p</i><0.01, **<i>p</i><0.05. Data points are reported as mean ± S.E.M. (n = 3) (<b>C</b>) Representative western blot of cells expressing αsyn-GFP<sub>11</sub>, treated with lactacystin (5 µM) for 24 hrs, using αsyn-specific antibody. (<b>D</b>) Western blots band quantification of cells expressing αsyn-GFP<sub>11</sub>. Bands were quantified by NIH ImageJ analysis software. GAPDH was used as loading control.</p

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