Fig. 7: The IL-17R–IκBζ axis facilitates restoration of Paneth cells from IFN-γ-induced damage.
a After treatment with IFN-γ (20 ng/ml) for 1 day, wild-type organoids were washed twice and then re-cultured in fresh media for 2 or 4 days in the absence or presence of IL-17A (20 ng/ml). Expression of the indicated genes was analyzed by RT-qPCR. The results are presented as the mean ± SD of triplicates and are representative of organoids from three mice. b Wild-type organoids treated as indicated were stained with UEA-1 and anti-E-cadherin antibody. Magnified images of the box are shown to the bottom. Results are representative of four independent experiments. The arrow indicates the release of Paneth cell granule into the crypt lumen. Scale bar, 50 μm. c Organoids from control (Nfkbizfl/fl) or Nfkbizfl/flVil1-Cre mice were treated with IFN-γ (20 ng/ml). The organoids were washed out, and re-cultured for 4 days in the absence or presence of IL-17A (20 ng/ml). Expression of the indicated genes was analyzed as in (a). d Wild-type organoids were treated as indicated, and analyzed by in situ hybridization (RNAscope) using probes specific to Nfkbiz, Enpep (an enterocyte marker), Lyz1 (a Paneth cell marker), and Lgr5 (an intestinal stem cell marker). Magnified images of the indicated boxes are shown to the bottom. Results are representative of two independent experiments. Scale bar, 20 μm (top) and 5 μm (bottom). The arrowheads in B and D indicate Paneth cells with the signals of Nfkbiz expression.
