Fig. 3: Inhibitory effect of EP on HMGB1 release and expression.

a, d Extracellular HMGB1 in the plasma of wild-type mice and lymphoma-bearing mice. b, e Extracellular HMGB1 in the plasma of lymphoma-bearing mice treated with 40 or 80 mg/kg EP. Mice injected with NaCl were used as control. Transferrin was used as loading control for plasma. c, f HMGB1 expression in lymphoma tissue. β-actin was used as loading control for cells. g, h Effect of EP on HMGB1 release in vitro determined by immunofluorescent microscopy (g) and western blotting (h). After cultured for 6 (Su-8) or 3 (Ly-3) h, DLBCL cells and medium were separately collected. After fix/permeabilization, cells were stained with rabbit anti-HMGB1 antibody, showing green color. DAPI staining (blue) indicates the nuclear localization. The green dots in the background indicate a typical phenomenon of HMGB1 release from the cells. Scale bar: 10 µm. Extracellular HMGB1 in the medium (M-HMGB1) and HMGB1 expression in cell lysate (C-HMGB1) were determined by western blotting. BSA and GAPDH were used as loading control. Numbers below each pair of bands are radios of interest proteins to the loading controls analyzed by densitometry. The significant differences (d–f) were analyzed by the student t-test