Fig 2.
Thoc1 gene deletion affects the histology of the small intestine but not that of the large intestine. (A) Representative H&E-stained small intestine tissue sections from tamoxifen-treated Thoc1F/F::Rosa26CreERT2 (Thoc1−) or Thoc1+/+::Rosa26CreERT2 (Thoc1+) mice are shown. Scale bars represent 200 μm. (B) Representative H&E-stained large intestine tissue sections from mice of the indicated genotypes treated as described for panel A. (C) Small intestine tissue sections from wild-type mice were immunostained for Thoc1 protein. A representative image is shown. The boxes represent the portion of the image magnified in the panels on the right to highlight staining of the crypt and villi. (D) The relative expression of Thoc1 protein in small intestinal villi (fractions V1 to V3, descending from villus tip) or crypts (fractions C1 and C2, descending to crypt bottom) isolated by differential tissue dissociation was assayed by Western blotting. Successful isolation of crypts and villi was verified by the presence of the crypt-specific marker CcnD1. Actin is the protein loading control. (E) Large intestine tissue sections from wild-type mice were immunostained for Thoc1 protein. A representative image is shown. The boxes represent the portion of the image magnified in the panels on the right to highlight staining of the crypt and differentiated epithelium. (F) The fraction of Thoc1 transcripts remaining at the indicated times from the start of tamoxifen treatment in the indicated tissues from Thoc1F/F::Rosa26CreERT2 mice relative to Thoc1+/+::Rosa26CreERT2 mice was quantitated by real-time RT-PCR. Each data point represents analysis of tissue from an individual mouse. The error bars represent the standard errors of the means. (G) Thoc1 protein levels in the small intestine of tamoxifen-treated Thoc1F/F::Rosa26CreERT2 (−) or Thoc1+/+::Rosa26CreERT2 (+) mice were assayed at the indicated times from the start of tamoxifen treatment by Western blotting. Actin is the protein loading control. (H) Thoc1 protein levels in the large intestine of tamoxifen-treated Thoc1F/F::Rosa26CreERT2 (−) or Thoc1+/+::Rosa26CreERT2 (+) mice were assayed as described for panel G.