Figure 1:
Methods for detection of potential HSF1 targets.
437 genes were selected for study based on either enrichment of their promoter in HSF1 chromatin IP as detected on a human promoter microarray containing 768 genes, presence of a sequence with high similarity to the previously described HSE in their promoter, or significant induction in heat shock microarray experiments. HSF1 binding was measured by Chromatin IP followed by quantitative PCR and expression by quantitative RT-PCR for each gene and this data was used to classify genes into HSF1-dependent and independent as well as heat-inducible and non-heat-inducible subsets. We calculated an optimized HSF1 binding motif based on the promoters that showed HSF1 binding. Finally, we tested promoter sequences of all classes for ability to confer heat-induced expression using a luciferase reporter system.
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