Fig. 7: IL-1R deficient EGCs impair tumor progression and SPP1+ TAM differentiation in colitis-associated CRC.
From: IL-1R signaling drives enteric glia-macrophage interactions in colorectal cancer
a Schematic representation of the murine AOM/DSS CRC model. Mice were intraperitoneally (i.p.) injected with azoxymethane (AOM, 10 mg/kg body weight) at day(d)0. Starting from d7, mice underwent 3 repetitive cycles of 1.5% dextran sodium sulfate (DSS) in drinking water as indicated. b GFAPCreAi14fl/fl mice were subjected to the AOM/DSS model (Fig. 7a) using 1% DSS per cycle. Representative images of EPCAM (white), IBA1 (green) and tdTomato (magenta) in tumor sections at d70 (scale bar 100 µm) (n = 3). c-e GFAPWtIL-1R1fl/fl and GFAPCreIL-1R1fl/fl mice were subjected to the AOM/DSS model (Fig. 7a). Tumors number and TAMs infiltration were assessed at d70. Tumor numbers of GFAPWtIL-1R1fl/fl and GFAPCreIL-1R1fl/fl littermates, representative images (left) and quantitative comparison of tumor numbers (right) (n = 14 mice per genotype) (c). Corresponding absolute numbers of SPP1+ and C1Q+ TAMs per mg tumor tissue (n = 7 GFAPWtIL-1R1fl/fl, n = 9 GFAPCreIL-1R1fl/fl) (d). Relative mRNA levels for Il6, normalized to the housekeeping gene Rpl32 in naive (n = 4) and AOM/DSS treated mice (n = 7 GFAPWtIL-1R1fl/fl, n = 5 GFAPCreIL-1R1fl/fl) (e). f GFAPCreAi14 fl/fl mice underwent the AOM/DSS model (Fig. 7a) using 1% DSS. Tumor or naive colon cells were isolated at d70 and FACS-sorted. Il6 expression levels of sorted tdTomatopos glial cells versus remaining tdTomatoneg cells of naive and AOM/DSS-treated GFAPCreAi14fl/fl mice (n = 4). Expression displayed as fold to Rpl32 and relative to naive tdTomatoneg cells. g SOX10CreERT2Ai14fl/fl mice were i.p. injected with Tamoxifen (1 mg in 100 µL sterile corn oil) on d–7, −6, and −5. Subsequently, mice were subjected to the AOM/DSS model (Fig. 7a) using 2% DSS per cycle. Representative image of tdTomato (magenta), IL-6 (green), and DAPI (blue) in tumor section at d70 (scale bar 100 µm) (n = 3). Data are represented as mean ± SEM (c–f). Statistical analysis: unpaired two-tailed t-test (c), unpaired two-tailed Mann-Whitney test (d), and two-way ANOVA with correction for multiple comparisons (e, f). *p < 0.05, ***p < 0.0005, ns not significant. Source data and exact p values are provided as a Source Data file.
