Fig. 3

MSI2 enhances HMGB1 production, nucleocytoplasmic translocation and extracellular release. A The protein expression of HMGB1 in SW620 and LOVO cells following 8 h of treatment with various concentrations of LPS. B-C HMGB1 protein expression in stable SW620 and LOVO cells cultured in the presence of LPS (10 μg/mL) for 2–8 h. D-E HMGB1 protein expression in stable SW620 and LOVO cells treated for 12 h with different concentrations of ABT-263 and ABT-737. F The concentrations of HMGB1 in the cytoplasm and supernatant of stable SW620 and LOVO cells were determined by ELISA. G Western blot analysis showed that loss of MSI2 impeded the translocation of ectopic HMGB1 into the cytoplasm and p65 into the nucleus in stable SW620 and LOVO cells. H Representative IFC images of stable SW620 and LOVO cells showing cytoplasmic translocation of ectopic HMGB1, the white arrows indicate cytoplasmic accumulation. Scale bars, 20 μm. I The nucleocytoplasmic transport pathway was significantly enriched among the upregulated KEGG pathways in MSI2-high patients in the TCGA CRC database. J The concentration of HMGB1 in the supernatant of stable SW620 and LOVO cells treated with LPS (10 μg/mL) or PBS for 8 h was determined by ELISA. K The release of HMGB1 into the extracellular supernatant of stable SW620 and LOVO cells treated with or without LPS (10 μg/mL) for 8 h was evaluated. These results are presented as the mean ± SD values; ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; A-F unpaired 2-tailed Student’s t test and J one-way ANOVA