Left: HMGB1 detection in HeLa cells by Immunocytochemistry. Cells were stained with mouse antibody to HMGB1 (red, 1:2,000 ), and co-stained with rabbit antibody to GAPDH (R-1701-100, green, 1:2,000). The HMGB1 antibody stains the chromatin binding protein HMGB1, which is localized in the nuclei. In contrast, the GAPDH antibody produces strong cytoplasmic staining. Right: Western blot analysis of HMGB1 expression (green) in cell lysates, using mouse antibody to HMGB1 at 1:2,000 dilution.  protein standard,  NIH-3T3,  C6,  HEK293,  HeLa, and  SH-SY5Y. The 25 kDa band corresponds to HMGB1 protein.