Fig. 6

HMGB1 induces tumor proliferation and PCNA expression through the ERK1/2 pathway. a, b Wild-type (WT) mice were induced CAC with AOM/DSS as described above, and in this process, they were intraperitoneally injected with or without neutralizing anti-HMGB1 antibody (HMGB1-ab) at days 1, 3, 5 during DSS treatment. Tumor tissues in the colon were excised at day 84 of the experimental procedure, and whole tissue extracts from the tumors were prepared and analyzed for PCNA expression by western blotting. a The representative images. b Quantitative analyses of proteins. Data are shown as means ± SEM from four mice in each group. Data shown are representative of three independent experiments. c CT26 cells (1 × 10³ cells/well) were seeded in 96-well plates. After 24 h, the cells were incubated for 1 h with or without U0126 (10 μM) before stimulation with HMGB1 (1 μg/ml) for 48 h, and then, the cell proliferation activity was measured by CCK-8 assay. Data are shown as means ± SD from five wells in each group. Data shown are representative of three independent experiments. d, e CT26 cells (1 × 10⁵ cells/well) were seeded in 6-well plates. After 24 h, the cells were incubated for 1 h with or without U0126 (10 μM) before stimulation with HMGB1 (1 μg/ml) for 48 h, and then, whole cell extracts were analyzed for PCNA expression by western blotting. d The representative images. e Quantitative analyses of proteins. Data are shown as means ± SEM from three independent experiments. NS not significant; *P < 0.05; **P < 0.01; ***P < 0.001