Figure 2

LPS or TNF-α induced HMGB1 cytoplasmic translocation in HepG2 cells. HepG2 cells were stimulated with LPS (100 ng/ml in panel A and B, 200 ng/ml in panel C) or TNF-α (25 ng/ml) for 20 h, and monitored for HMGB1 cytoplasmic translocation by immunocytochemistry (Panel A) or by Western blot analysis after cell fractionation (Panel C). A, B). HMGB1 immunohistochemistry assay. The relative fluorescence intensity in the nuclear ("N") or cytoplasmic ("C") regions of multiple represent cells were determined using the Image Proplus Software, and expressed as mean ± SEM (in arbitrary units, AU) of three independent experiments. Red: nuclear; green: HMGB1; yellow: merge (original magnification × 400). * P < 0.05 versus control. C). HMGB1 Western blotting analysis. Following cell fractionation, HMGB1 content in the cytoplasmic ("C") or nuclear ("N") fraction was determined by Western blot analysis.