Fig 2.

Transcriptional activation of HSP70B promoter by HSF1 in normal (RSK2+/+) and CLS (RSK2−/−) cells at 37°C. Wild-type HSF1 expression plasmid (pcDNA3.1HSF1; 0.2 μg/well) or empty control (pcDNA3.1; 0.2 μg/well) were cotransfected with HSP70B promoter reporter construct (HSP70B-Luc) (0.6 μg/well) along with pSV-β-galactosidase plasmids (0.6 μg/well), respectively, transfected into normal (RSK2+/+) and CLS (RSK2−/−) cells in triplicate and incubated for 20 to 24 h. Wild-type HSF-1 were force-cloned into XhoI/EcoRI sites of the pcDNA3.1 vector as described (Cahill et al 1996). HSP70B-Luc reporter plasmid was constructed as described before (Chen et al 1997). Cells were harvested for luciferase assay. Luciferase activity was normalized to β-galactosidase activity. Luciferase activity is expressed as fold activity over the empty pcDNA3.1 plasmid control