Fig. 2

DOE inhibits LPS-triggered release of HMGB1 in RAW 264.7 cells. a Cells cultured for 24 h in serum-free medium were triggered by vehicle (DMSO) or LPS in the presence or absence of DOE. Following incubation for 24 h, aliquots of conditioned media or whole-cell lysates were analyzed by immunoblot analysis. Ponceau S staining and β-actin were used as the loading controls. b Cells were treated with vehicle (DMSO) or LPS in the presence or absence of DOE for 24 h, and then whole-cell lysates were fractionated into cytosolic (C) and nuclear (N) fractions. The localization of HMGB1 was analyzed by immunoblotting with the indicated antibodies. Representative blots from three independent experiments are shown