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. 2009 Oct 29;28(23):3758–3770. doi: 10.1038/emboj.2009.298

Figure 2.

Figure 2

Effect of Hsp70 on the morphology and cytotoxicity of αSyn aggregating species throughout the reaction. (A) Fibril formation by αSyn was monitored by ThT fluorescence. Samples correspond to untreated or Hsp70-treated αSyn at a 1:10 ratio, in the absence or presence of ATP. Samples labels in (B). A negative control with added ovalbumin was used. AU, arbitrary units. (B) TEM analysis of samples corresponding to 24 and 64 h of the aggregation reaction. A negative control with ovalbumin was also included. Scale bar, 500 nm. (C) The toxicity levels of the soluble species from samples corresponding to the different conditions and time points of the reaction were determined by LDH release measurement (left axes) using human SH-SY5Y cells. Samples correspond to untreated αSyn (white bars) or αSyn treated with 1:10 Hsp70:αSyn (black bars), either in the absence (left panel) or presence (right panel) of ATP. The bars represent the intrinsic toxicity, that is the net toxicity (with background toxicity levels subtracted) normalized to the αSyn concentration. In all cases, values represent the average results of duplicate experiments. Connected circles represent the Hsp70 soluble levels at each time point (right axis).