FIGURE 2.

Protein kinase A is required for ethanol-mediated increases in TH protein levels. A, cells were treated without or with 100 mm ethanol for 24 h, to which kinase inhibitors were added for the last 9 h of the ethanol treatment as indicated: 1 μm Bis, 1 μm PP2, 5 μm H89, and 40 μm Rp-cAMPS. TH protein levels were detected by Western blot analysis. The image is representative of six experiments. B, cells were treated without (con) or with 100 mm ethanol for 24 h alone (Et) or in combination with 1 μm H89 for the last 9 h (Et/H89). TH was immunoprecipitated using anti-TH antibody and immunoblotted with anti-phospho(Ser/Thr) PKA substrate antibody. The image is representative of four experiments. C, cells were treated without or with 100 mm ethanol for 24 h, to which 1 μm H89 was added for the last 9 h. Phosphorylation of TH was detected by Western blot analysis with anti-phospho(Ser-40)TH antibodies. The image is representative of three experiments.