Skip to main content
. 2004 Feb 11;101(8):2452–2457. doi: 10.1073/pnas.0306734101

Fig. 2.

Fig. 2.

Conditional intestinal epithelial cell-specific IKKβ knockout mice. (A) Intestine-specific expression of Cre recombinase mRNA in Vil-Cre mice. mRNA expression of Cre and β-actin was assessed by RT-PCR of RNA extracted from the indicated tissues of Vil-Cre mice. SI, small intestine. (B) Intestinal epithelial cell-specific recombination of loxP sites in Vil-Cre mice. β-Galactosidase activity was assessed in the small intestine of Cre/loxP recombination reporter mice (Gtrosa26) and Gtrosa26/Vil-Cre mice by staining with X-Gal and nuclear fast red (counterstain). Blue staining indicates loxP site recombination in Gtrosa26/Vil-Cre epithelial cells of crypts and villi. (C) Schematic representation of IkkβF and IkkβΔ alleles. H3, HindIII site; a, b, c, and d, annealing sites of primers used for genotyping (see supporting information); E3, exon 3; triangles, loxP sites; Probe, annealing site for Southern blot probe. (D) Southern blot analysis of the Ikkβ locus using HindIII-digested DNA from isolated intestinal epithelial cells of IkkβF/Δ and Vil-Cre/IkkβF/Δ mice. The positions of the IkkβF and IkkβΔ restriction fragments are indicated. (E) Defective IR-induced IKK and NF-κB activation in IKKβ-deficient intestinal epithelial cells. Intestinal epithelial cells were isolated from IkkβF/Δ and Vil-Cre/IkkβF/Δ mice at the indicated times after 8-Gy IR. IKK activity was determined by an in vitro kinase assay (KA), and IKKα levels (loading control) were assessed by Western blotting (WB). NF-κB binding activity was determined by electrophoretic mobility-shift assay (EMSA). Fold induction by IR of IKK and NF-κB binding activities relative to epithelial cells from nonirradiated control mice is shown under each lane. (F) Normal intestinal epithelial lineage development in the absence of IKKβ. Sections of the small intestine from 6-week-old IkkβF/Δ and Vil-Cre/IkkβF/Δ mice were prepared and stained as indicated from left to right to detect enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. H&E, hematoxylin and eosin.