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. 2000 Nov 21;97(25):13967–13972. doi: 10.1073/pnas.240452197

Figure 6.

Figure 6

CREB2 activates Gadd153 transcription through the GABAB receptor. (a) Dose-response to baclofen. Transfected CHO-GABAB cells were treated with baclofen (0.1 μM-1 mM; 6 h) and cytoplasmic extracts were prepared. CAT levels, from the Gadd153-CAT reporter gene (14), were determined by ELISA assay. Baclofen induced CAT expression in a dose-dependent manner with an EC50 of 4 μM. (b) Effects of CGP54626, PTX, and PD98059 on baclofen-induced CAT production. Cells were pretreated with CGP54626 (1 μM; 30 min), PTX (50 ng/ml; 16 h), or PD98059 (50 μM; 30 min) and CAT levels were determined in cytoplasmic extracts after baclofen treatment (100 μM; 6 h). All three compounds blocked baclofen/CREB2-mediated CAT expression. Transfection of cells with vector (pcDNA3) instead of CREB2 greatly diminished CAT expression levels, and resulted in a markedly decreased response to baclofen.