(A) Illustration of β-catenin/Ctnnb1 conditional knock-out (CKO) in Tert+ cells (TERTTCE/+;Ctnnb1fl/fl).
(B) Scheme of mouse treatment.
(C) β-catenin CKO-induced downregulation of Wnt/β-catenin target gene expression in Tert+ cells. qRT-PCR of Tert+ cells for CD44, CD133, and Axin2 expression from TertTCE/+ (control) and TertTCE/+:Ctnnb1Δ/Δ (experimental group) treated with tamoxifen.
(D–K) IHC of impaired intestinal regeneration by β-catenin CKO in Tert+ cells. H&E staining (D); CK19 (E); Ki67 (F); CC3 (G); lysozyme (H); PAS (I); villin (J); ChgA (K). Scale bars=20μm.
(L–M) Cell cycle-related gene expression in Tert+/+;Ctnnb1+/+ and TertTCE/+;Ctnnb1Δ/Δ (10 Gy, 7 dpi). Cyclin D1 (L); c-Myc (M). Scale bars=20μm. Cyclin D1 and c-Myc were significantly downregulated by β-catenin CKO in Tert+ cells and WBI.
(N) No loss of Tert+ cells by β-catenin CKO during intestinal homeostasis. Population analysis of Tert+ cells after Tam treatment in TertTCE/+ and TertTCE/+;Ctnnb1Δ/Δ, using FACS. ns: non-significant (p>0.05).
(O) Reduced proliferation of Tert+ cells during intestinal regeneration by β-catenin CKO. Quantification of Tert+:Ki67+ cells of Tam and IR (10 Gy; 2 dpi)-treated TertTCE/+ and TertTCE/+;Ctnnb1Δ/Δ mice, using FACS.
The representative images are shown; N≥3; error bars indicate s.e.m.