Fig 2. APC-mutant intestinal epithelial cells are defective in IL-22 mediated gene regulation.

(A). WT and Apc^Min/Min organoids were treated with IL-22 (2 ng/ml) for 3 hours. RNA was isolated for RNAseq analysis. Graph shows the fold change of genes induced by IL-22 in WT and Apc^Min/Min organoids. The 11 genes shown were the only genes up-regulated by IL-22 in Apc^Min/Min organoids (p < 0.1). (B) Expression of candidate up-regulated genes from RNAseq data were verified by RT-qPCR. Data shown are relative levels of mRNA for genes of interest relative to Tbp expression. At least 3 independent experiments were performed in each case. *p < 0.05, **p < 0.01, ***p < 0.001 on paired t test. (C). WT and Apc^Min/Min organoids were treated with IL-22 (10 ng/ml) for 3, 6, or 24 hours, and RT-qPCR analysis was performed. Data show the relative expression of mRNA for Reg3g or Socs3 compared to Tbp. Three independent experiments were performed. *p < 0.05, **p < 0.01, paired t test. Numerical values for (A), (B), and (C) are available in S1 Data. APC, adenomatous polyposis coli; IL-22, interleukin-22; Reg3g, regenerated islet-derived protein 3 gamma; RNAseq, RNA sequencing; RT-qPCR, quantitative reverse transcription polymerase chain reaction; Socs3, Suppressor of cytokine signalling 3; Tbp, TATA box binding protein; WT, wild-type.