FIGURE 5.

Chronic ethanol induces an association of TH with heat shock protein 90. A and B, cells were treated without (Con) or with 100 mm ethanol for 24 h (Et), or with 100 mm ethanol for 24 h to which 1 μm GA was added for the last 9 h (Et/GA). TH was co-immunoprecipitated with HSP90 using anti-HSP90 antibody (A), and HSP90 was co-immunoprecipitated with TH using anti-TH antibody (B). HSP90 and TH levels in input samples were analyzed by Western blot. Images are representative of four experiments. C, cells were treated without (Con) or with 100 mm ethanol for 24 h (Et). Akt was co-immunoprecipitated with HSP90 using anti-HSP90 antibody (upper panel), and HSP90 was co-immunoprecipitated with Akt using anti-Akt antibody (lower panel). Images are representative of three experiments. D, cells were treated without (Con) or with 100 mm ethanol (Et) for 24 h alone or together with 5 μm H89 for the last 9 h (Et/H89). Cells were homogenized for co-immunoprecipitation of HSP90 with anti-TH antibody. The image is representative of three experiments. E, cells were treated without or with 100 mm ethanol for 24 h, to which 1 μm H89 were added for the last 9 h of the ethanol treatment. HSP90 protein levels were detected by Western blot analysis. The image is representative of six experiments.