Fig. 2.

Sublethal heat shock does not increase clusterin levels in motor neurons or astrocytes, or generally in spinal cord–DRG cultures. a Immunolabeling of cytoplasmic clusterin in motor neurons and astrocytes using antibody SC-6419. Cultures were maintained at 37 °C for 24 or 48 h after heat shock at 43 °C for 30 min. Graphs show quantitation of clusterin levels in each cell type as mean pixel density of fluorescence of cy3-anti-goat IgG. Shown are means ± SEM, three cultures per condition and minimum of 20 cells per culture. Levels of clusterin in whole cultures were measured by SDS-PAGE/Western analysis, using stress inducible Hsp70 (HspA1) as a positive control for heat shock and Hsc70 (HspA8) as a negative/loading control. b Experiments conducted using antibody to nuclear clusterin (SC-8354) showing no change in expression with heat shock (p > 0.05, one-way ANOVA)