Supplementary Figure 5.

EGFR signaling in myeloid cells protects from colitis. (A) Scheme of the DSS-dependent colitis model. On day 10, mice were starved for 4 hours before oral administration of FITC-dextran. (B) BrdU IF staining, arrowheads depict BrdU⁺ IECs. Scale bars 50 μm. (C) Quantification of cleaved Caspase-3⁺ IECs per crypt as readout for apoptosis (30 crypts per mouse were counted, n = 3–4). (D–H) Flow cytometric analysis of the IEL and LP immune cell fraction of colons from DSS-treated mice shows percentage of live cells of (D) TCRβ⁺ αβ T cells, (E) ratio of CD4⁺/CD8⁺ T cells, (F) CD19⁺ B cells, (G) CD11c⁺/CD11b⁺/CD103⁺, and (H) CD11c⁺/CD11b^-/CD103⁺ dendritic cell subsets. (I) Quantitative reserve transcriptase PCR analysis of IL6, CXCL1, CXCL2, tumor necrosis factor-α, and IL1β mRNA expression. Relative mRNA expression levels were determined in whole colon tissues on day 10 (n = 3) and normalized to Cyclophilin. Data are mean ± SEM. (J–K) IL6 serum levels in endpoint mice with (J) AOM/DSS treatment (n = 4 for Egfr^wa/wa, otherwise n = 7) and (K) the Apc^Min background (n ≥7). Data are mean ± SEM. #P < .5, 1-way analysis of variance with Dunn’s posttest.