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. 2000 Oct 15;20(20):7510–7516. doi: 10.1523/JNEUROSCI.20-20-07510.2000

Fig. 2.

Fig. 2.

Localization of the IL-1/TNF response element of the ACT gene. A, Structure of the DNA fragment containing the ACT gene and its 5′ flanking region (GenBank accession no. AL049839). Exons coding for protein C inhibitor (PCI) and antichymotrypsin (ACT) are indicated by numbers.B marks restriction sites for BamHI.Asterisk indicates the end of the DNA fragment harbored by phage. BamHI–BamHI fragments used for construction of reporter plasmids are marked by letters (a–g). B, Human astrocytes were transfected with plasmids pact-3573CAT, ptkCATDEH, p'd'ACTtkCAT, p'e'ACTtkCAT, p'f'ACTtkCAT, or p'g'ACTtkCAT, and β-galactosidase expression vector as internal control for transfection efficiency. One day after transfection, cells were stimulated with the indicated cytokines, cultured for another 24 hr, and harvested. CAT activities were normalized to β-galactosidase activities (cpm/unit × 10−3). tkindicates thymidine kinase minimal promoter from ptkCATDEH.C, Human astrocytes were transfected with pStACTCAT, p'a'StCAT, p'b'StCAT, p'c'StCAT, p'd'StCAT, p'e'StCAT, p'f'StCAT, p'g'StCAT, and pCH110. Cells were treated with cytokines and harvested as described above. CAT activities were normalized to β-galactosidase activities (cpm/unit × 10−3). OP indicates 244-bp-long ACT promoter containing elements mediating response to OSM but not to IL-1 or TNF.