Figure 2.

Ablation of Rnf43/Znrf3 leads to a wrinkled tongue and thinner tongue epithelium

(A) Schematic illustration of the experimental design. Control (Rnf43^fl/fl;Znrf3^fl/fl) and Rnf43^fl/fl;Znrf3^fl/fl;Krt5-Cre^ERT2 mice were treated with tamoxifen for five continuous days (D0–D4), and tongue tissue was collected at day 7. TMX, tamoxifen.

(B) Representative bright-field images of tongues collected from control (n = 3) and RZ dKO (n = 3) mice.

(C) Representative images of hematoxylin and eosin (H&E) staining of anterior tongue sections from control and RZ dKO mice. Note the thinner lingual epithelium (arrows) in RZ dKO mice. n = 3 for each group. Scale bars, 50 μm.

(D) qRT-PCR analysis of the expression of Rnf43 and Znrf3 in tongue epithelium in control and RZ dKO mice. Expression levels of Rnf43 and Znrf3 were normalized to Gapdh. Data are presented as mean ± SEM. ^∗∗∗p < 0.0001. n = 3 for control group and n = 4 for RZ dKO group.

(E) Immunofluorescence staining of sections of anterior tongues of control and RZ dKO mice for KI-67 (red) and E-cadherin (ECAD, green). We frequently noted a single layer of sparse KI-67⁺ cells in the dorsal lingual epithelium in RZ dKO sections (arrow), but not in sections from control mice. Dashed circles show fungiform papillae. Cell nuclei were counterstained with DAPI (blue). n = 3 for each group. Scale bars, 50 μm.

(F) Tabulation of KI-67⁺ cells in the dorsal lingual epithelium. Fewer KI-67⁺ cells are present in the dorsal lingual epithelium of RZ dKO mice than of control mice. Data are presented as mean ± SEM. ^∗∗∗p = 0.0005. n = 3 for each group. See also Figures S1.