After 40 min at 43uC (HS) or at the indicated times during recovery at 37uC (recovery), recruitment of HSF1 to the COX-2 and HSP70 promoters was analyzed by ChIP assay. (C) ChIP- enriched DNAs using preimmune serum (IP NS IgG) or anti- HSF1 serum (IP anti-HSF1), as well as input DNAs (INPUT) were prepared, and DNA fragments of the COX-2 gene (22629 to 22420) and HSP70 gene (2262 to 270) were amplified by PCR. (D) Quantification of ChIP assay shown in (C). Samples from at least three independent experiments were analyzed by real time PCR. Relative promoter occupancy is expressed as fold induction of control arbitrarily set to a value of 1. Error bars indicate 6 S.D. (TIF) Figure S3 Stable suppression of HSF1 by RNA interfer- ence reduces heat-induced COX-2 mRNA expression in HCT116 cells. pHCT116-retro and pHCT116-HSFi cells were subjected to heat shock at 43uC or left untreated (control). 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