Figure 3

Stimulated macrophages demonstrated a nuclear retention of HMGB1 in co-cultures with oxaliplatin. RAW 264.7 macrophage-like cells were cultured without exogenous stimulus (a) or with IFN-γ and LPS alone (b) or with IFN-γ and LPS in the presence of 0.5 μM oxaliplatin (c). Immunostaining for HMGB1 (green), nucleus (blue) and the cytoskeletal protein β-actin (red) was performed in fixed cells after a culture time of 24 h. Almost all nuclei demonstrated strong HMGB1 expression in resting cells (a), while the nuclear HMGB1 was as expected substantially reduced in IFN-γ- and LPS-activated macrophages (b). Macrophages that had been activated in the same way in the presence of oxaliplatin demonstrated a strong nuclear retention of HMGB1 (c).