Fig. 2.

FRT only affects recruitment of NFκB-p65 to the −586/−468 κB enhancer site in the TNF-α promoter. RAW 264.7 macrophages were prewarmed to 37°C or 39.5°C for 30 min, stimulated with 100 ng/ml LPS and incubation was continued at the same temperature for an additional 30 min. Cross-linked chromatin was immunoprecipitated with anti-p65 antibody and analyzed for recruitment of NFκB-p65 to the κB binding sites by qRT-PCR using primer pairs spanning the regions −364/−182 (a), −586/−468 (b), −685/−543 (c), and −912/−763 (d) and expressed as fold-change vs. unstimulated 37°C cells. Data presented as mean ± SE of four independent experiments. *p < 0.05 compared with untreated controls. †p < 0.05 vs. 37°C cells at the same time point