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. Author manuscript; available in PMC: 2012 Apr 1.
Published in final edited form as: Cytokine. 2011 Jan 26;54(1):61–67. doi: 10.1016/j.cyto.2010.12.017

1. Exposing A549 cells to HS recruits HSF1 to the HSP72 gene and induces HSP72 mRNA and protein expression.

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A. A549 cells were heat-shocked at 42°C for 1h, DNA and protein were cross-linked with formaldehyde, and HSF1 recruitment to the HSP72 gene analyzed by ChIP assay. B. For analysis of HSP72 mRNA levels, A549 cells were heat-shocked at 42°C for 2h then switched to 37°C. Total RNA was prepared either immediately after HS or after 1, 2 or 4h recovery at 37°C and analyzed by qRT-PCR for HSP72 and standardized for GAPDH. Data are presented as mean±SE (n=4), * denotes p<0.05, in comparison to non-heat shocked controls. C. For analysis of HSP72 protein levels, cells were heat shocked at 42°C for 2h then switched to 37°C for 4h. Cells were lysed and HSP72 protein was analyzed by immunoblot and standardized to β-tubulin. Figure representative of two independent experiments.