Journal: PLoS ONE
Article Title: Elevation in Body Temperature to Fever Range Enhances and Prolongs Subsequent Responsiveness of Macrophages to Endotoxin Challenge
Figure Lengend Snippet: HSP70 plays a role in mediating thermally-enhanced TNF-α production in macrophages. A , Peritoneal macrophages were isolated from LPS-challenged mice after 2 hour heat treatment. Cells were recovered overnight and re-stimulated (1×10 6 /well) in vitro with LPS and IFN-γ at 37°C for 4 hours, then HSP70 mRNA level was measured by quantitative real-time PCR. The results are presented relative to GAPDH and baseline expression in unstimulated cells from RT-mice. B , Macrophages (2×10 5 /well) from LPS-challenged mice were re-stimulated with LPS and IFN-γ at 37°C for 2, 6 or 24 hours to examine HSP70 secretion by ELISA. C , Macrophages (2×10 5 /well) from LPS-challenged mice were re-stimulated with LPS and IFN-γ at 37°C for 6 hours with or without HSP70 inhibitors: KNK437 (20, 10 µM) or Pifithrin-µ (5, 1 µM) to detect TNF-α production by ELISA. Cells from each treatment condition were pooled from 2–4 mice and measured in triplicate. Data are mean ± SD. Data are representative of two independent experiments. * In comparison of cells with and without HSP70 inhibitors from WBH-mice. ** In comparison of cells with and without HSP70 inhibitors from RT-mice. *, ** p
Article Snippet: In some experiments, macrophages were treated with HSP70 inhibitors (KNK437, EMD chemicals; Pifithrin-µ, Sigma-Aldrich) together with 100 ng/mL LPS and 25 U IFN-γ.
Techniques: Isolation, Mouse Assay, In Vitro, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay