Figure 1

LPS and TNF-α stimulated HepG2 cells to release HMGB1 in a time- and dose-dependent fashion. A, B) LPS and TNF induced HMGB1 release in a time-dependent fashion. HepG2 cells were stimulated with LPS (200 ng/ml) or TNF-α (25 ng/ml) for indicated time periods, and extracellular HMGB1 levels (Panel A) or cell viability (Panel B) or were determined by MTT or Western blotting analysis, respectively. HMGB1 levels were determined by the relative optical intensity (OD) of the immunoreactive bands on Western blots, and expressed as mean ± SEM of three experiments in duplicate. * P < 0.05 versus 0 h treatment group. D, E, F, G) LPS and TNF induced HMGB1 release in a dose-dependent manner. HepG2 cells were stimulated with LPS or TNF-α at indicated doses for 24 h cells viability and extracellular HMGB1 levels were determined as described above. *P < 0.05 versus control ("-LPS", Panel C, D; or "-TNF", Panel E, F).