FIGURE 2.
Overexpression of Hsp72 renders macrophages resistant to H2O2-induced HMGB1 release. A, Western blotting analysis of Hsp72 levels in RAW 264.7 cells transfected with empty plasmid (lane 1), Hsp72 expression construct (lane 2) or challenged with mild heat shock (HS, 42.5°C, 1 h). Tubulin was used as a loading control. B, Visualization of fluorescent Hsp72 protein in RAW 264.7 cells transfected with Hsp72 expression construct (top), or empty plasmid (bottom). Nuclei were visualized by Hoechst staining. C, Western blotting analysis of H2O2-induced HMGB1 release in RAW 264.7 cells transfected with empty plasmid, or Hsp72 expression construct. RAW 264.7 cells were stimulated with H2O2 (0.125 and 0.25 mM) for 12 h, and HMGB1 levels in the culture medium were determined, and expressed as mean ± SEM of three experiments in duplicate. *, p < 0.05.