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Fig. 6. The proximal 5-aminolevulinic acid synthase 1 (ALAS1) promoter activates basal transcription, but does not mediate drug response. Two fragments of the human ALAS1 promoter fragment were amplified and cloned into the pGL3tk luciferase reporter vector. The core promoter was contained in a 234-bp fragment, which ranges from –234 bp to the start site of the human ALAS1 transcript. A longer construct, consisting of the sequence from –1,249 bp to the transcriptional start site also was tested. These constructs were transiently transfected into leghorn male hepatoma (LMH) cells. At 4 h after transfection, cells were exposed to either vehicle only (0.1% DMSO) or phenobarbital (PB; 400 m M), Rifampicin (Rif; 10 m M), or propylisopropylacetamid (PIA, 250 m M) for 24 h. Luciferase activity was measured in cell lysates and normalized against b-galactosidase levels. Luciferase levels are expressed relative to the levels of vehicle-treated cells. Data represent the mean of three independent experiments plus one SD.