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. 2021 Jan 22;10:e59459. doi: 10.7554/eLife.59459

Figure 5. Loss of Wnt5a results in loss of Gli1+ mesenchymal stem cells (MSCs).

(A and B) Ki67 staining (red) of incisors from Gli1-LacZ (control) and Axin2-CreERT2;Wnt5afl/fl;Gli1-LacZ mice 3 weeks after tamoxifen induction. (C) Quantitation of the percentage of Ki67+ transit amplifying cells (TACs) from (A and B). (D–G) β-gal staining (green) of incisors from 1-month-old Gli1-LacZ and Axin2-CreERT2;Wnt5afl/fl;Gli1-LacZ mice 3 weeks after tamoxifen induction. Boxes in (D and E) are shown magnified in (F and G), respectively. (H) Quantitation of the percentage of Gli1+ cells per higher magnification section (F and G) of Gli1-LacZ and Axin2-CreERT2;Wnt5afl/fl;Gli1-LacZ incisor mesenchyme. Schematic at the bottom indicates induction protocol. The white dashed lines outline the cervical loop. Arrows indicate positive signal and asterisks indicate diminished signal. All quantitative data are presented as mean ± SD. ns, no significance. ****, p<0.0001. Four mice with four sections within each mouse per group were used to quantify Ki67+ cells. Gli1+ cells in the proximal region between the two cervical loops were counted in the mouse incisor. Scale bars, 100 μm.

Figure 5—source data 1. Source data for Figure 5C and H.

Figure 5.

Figure 5—figure supplement 1. Validation of Wnt5a knockout efficiency in Axin2-CreERT2;Wnt5afl/fl incisors.

Figure 5—figure supplement 1.

(A–D) RNAscope (red) of Wnt5a in incisors from Wnt5afl/fl (control) and Axin2-CreERT2;Wnt5afl/fl incisors 3 days after tamoxifen induction. White arrow indicates positive signal and asterisk indicates diminished signal. Schematic at the bottom indicates induction protocol. Scale bars, 100 μm.
Figure 5—figure supplement 2. Loss of Wnt5a results in diminished EdU+ label retaining cells (LRCs).

Figure 5—figure supplement 2.

(A–D) EdU staining of Wnt5afl/fl and Axin2-CreERT2;Wnt5afl/fl mouse incisors. (E) Quantification was performed by calculating the percentage of EdU+ cells per section under higher magnification (B and D) in Wnt5afl/fl and Axin2-CreERT2;Wnt5afl/fl mouse incisor mesenchyme. Schematic underneath figures indicates induction protocol. All quantitative data are presented as mean ± SD. ***, p<0.001. Scale bars, 100 μm.
Figure 5—figure supplement 2—source data 1. Source data for Figure 5—figure supplement 2E.
Figure 5—figure supplement 3. Loss of Wnt5a results in thicker dentin and diminished transit amplifying cells (TACs).

Figure 5—figure supplement 3.

(A–D) H and E and Ki67 staining (red) of incisors from 1-month-old Wnt5afl/fl (control) and Axin2-CreERT2;Wnt5afl/fl mice 6 weeks after tamoxifen induction. (E–H) H and E and Ki67 staining (red) of incisors from Wnt5afl/fl (control) and Axin2-CreERT2;Wnt5afl/fl mice 5 months after tamoxifen induction. Black arrows in A and E indicate normal dentin and black arrows in B and F indicate thicker dentin. White arrows in C and G indicate positive signal and asterisks in D and H indicate diminished signal. White dashed lines outline the cervical loop. Schematics at the bottom indicate induction protocols. Scale bars, 100 μm.