Fig. 2
MSI2 modulates tumor immunity in CRC via HMGB1-mediated DC maturation and migration. A Enrichment analysis of the differentially expressed proteins showed that MSI2 regulated HMGB1 expression in stable LOVO, SW620, and HT29 cells. Red arrows, MSI2; blue arrows, HMGB1. B Western blot analysis of MSI2 and HMGB1 expression in stable LOVO, SW620, and HT29 cells. C FACS analysis of the DC maturation markers CD80, CD86, CD40, and CD83 after coculture of iDCs with stable LOVO cells and rh-HMGB1 for 24 h. D Statistical analysis of the expression of the DC maturation markers CD80, CD86, CD40, and CD83. E HMGB1 mRNA expression was determined by qRT–PCR in untreated and CAC mice, n = 5. F Western blot analysis of MSI2 and HMGB1 expression in colonic mucosa from untreated and CAC mice. G-H FACS and statistical analysis of DCs (CD11b+CD11c+MHCII+) isolated from the SP, MLNs, and CLP of untreated (n = 4) and CAC (n = 5) mice. I-J FACS and statistical analysis of CD4+ T and CD8+ T-cell percentages among cells isolated from the SP, MLNs, and CLP of untreated (n = 4) and CAC (n = 5) mice. K Representative IHC images of MSI2, HMGB1, CD11c and CD3 staining in CAC mice. L Statistical analysis for MSI2, HMGB1, CD3 and CD11c average IHC staining density (n = 5). Scale bars, 100 μm. M FACS analysis of CD103 expression with gating on DCs (CD11b+CD11c+MHCII+) in cells isolated from MLNs and CLP of untreated and CAC mice. N Statistical analysis of the CD103 mean fluorescence intensity (MFI) in DCs isolated from the SP, MLNs, and CLP of untreated (n = 4) and CAC (n = 5) mice. O-P Representative IFC images and statistical analysis of CD11c-positive cells in Msi2Transgenic and WT CAC mice. Scale bars, 100 μm. These results are presented as the mean ± SD values; ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; D one-way ANOVA, L Mann-Whitney test and E, H, J, N, P unpaired 2-tailed Student’s t test