Figure 2

Protein inclusions remain insoluble in lysis buffer and FloIT analyses can be performed up to several hours post-lysis.

(a) Time series of confocal microscopy images showing cells expressing SOD1^G93A-eGFP imaged every 6 s following the addition of lysis buffer. The release of soluble SOD1^G93A from the newly lysed cell is visible at 30 s while the inclusions remained unchanged for at least 10 minutes. Scale bar (bottom right) is 5 μm. (b) N2a cells transfected to express the indicated protein were lysed at a density of 150,000 cells/ml. The lysates were kept on ice over the entire 3 h time course and were mixed briefly before aliquots were taken for FloIT measurement at the times indicated. Data are plotted as % of t = 0 values and are means ± SEM (n = 3). Results shown are each representative of two independent experiments.