Fig. 5

DOE inhibits LPS-triggered release of HMGB1 through inhibition of NO formation in RAW 264.7 cells. a and b Cells cultured for 24 h in serum-free medium were pretreated with or without L-NMMA and then stimulated with vehicle (DMSO) or LPS in the presence or absence of DOE. After incubation for 24 h, aliquots of whole-cell lysates or conditioned media were subjected to immunoblot or nitrite analysis, respectively (s). The levels of HMGB1 were analyzed by immunoblot using equal volumes of conditioned media (b). Ponceau S staining and β-actin were used as the loading controls. The blots are representative of three independent experiments. The results are plotted as the means ± S.E. (n = 6). ^*, p < 0.01 vs untreated group; ^#, p < 0.01 vs LPS-treated group