Fig. 4: Immunocompetence-dependent tumor-promoting and -inhibiting function of MEN1 in colon cancer.

a, Cell proliferation rate of HCT116 in 2D cell culture with and without knockout of MEN1. NS, not significant. Each data point represents mean ± s.e.m. cell counts (n = 3 for each arm). Two-way ANOVA was used for statistical analysis. b, HCT116 xenograft tumor growth rate in immunodeficient mice with and without knockout of MEN1. Each data point represents mean ± s.e.m. tumor volumes (n = 5 for each arm). Two-way ANOVA was used for statistical analysis. *P < 0.05, **P < 0.01. c, Schematic view of murine CT26 engraftment experiment design. d, Tumor growth in immunodeficient NSG mice inoculated with control (sgEV) or Men1 knockout (sgMen1-1, sgMen1-2) CT26 cells. Each data point represents mean ± s.e.m. tumor volumes (n = 12 for each arm). **P < 0.01. e, Tumor growth in immunocompetent BALB/c mice inoculated with control (sgEV) or Men1 knockout (sgMen1-1, sgMen1-2) CT26 cells. Each data point represents mean ± s.e.m. tumor volumes (n = 5 for each arm). Two-way ANOVA was used for statistical analysis. ***P < 0.001. f, Heatmap showing expression value (z-score based on DESeq normalized RNA-seq counts) of differential genes from Men1 knockout versus control tumors in immunocompetent mice. Two control samples and four Men1 knockout tumors were subjected to RNA-seq analysis. g, GO analysis of differentially expressed genes in Men1 knockout versus control cells was performed and the top five terms are shown. The x axis represents the number of genes. Wald tests defined in DEseq2 were used to calculate P values. h,i, Quantification (h) and representative immunofluorescence images (i) of control (sgCtrl) or Men1 knockout (KO) CT26 cells for the detection of dsRNA. Red signal, dsRNA (J2 antibody); blue signal, DAPI. Scale bars, 20 μm. Each bar in the left-hand panel represents the mean of quantifications from 30 randomly picked fields (unpaired two-tailed Student’s t-test). **P < 0.01, ***P < 0.001. CTCF, corrected total cell fluorescence.