Figure 8.

Overexpression of HSP60 increases neurotoxicity of HEK293 cell lysates added to cocultures of neurons and microglia. HEK293 cells were transfected with plasmids encoding hHSP60 or GFP (control). A, Immunoblot of lysed cells probed with the SPA-807 antibody directed against HSP60. Left lane, Control-transfected; right lane, HSP60-transfected. β-Actin served as a loading control. B, Quantitation of NeuN-positive neurons as a percentage of all DAPI-positive cells in cocultures of neurons and microglia derived from wild-type mice treated with cell lysates of hHSP60- or control-transfected HEK293 cells. Cell lysates were diluted 1:15. Quantitation of NeuN-positive neurons was performed by analyzing four high-power fields per coverslip. For each condition, experiments were performed in duplicates. The data shown are pooled from three individual experiments with three different preparations of transfected cells. The results are presented as mean ± SD (*p < 0.001) for the comparison of control-transfected cells with hHSP60-transfected cells (one-way ANOVA and post hoc Bonferroni's test).