Fig. 6: Canonical Wnt signaling prevents premature differentiation of CXCL12+ AP cells.
From: Wnt-directed CXCL12-expressing apical papilla progenitor cells drive tooth root formation
Identification of mechanisms underlying canonical Wnt inactivation-induced CXCL12+ AP cell fate shifts by RNA-req. a, b Workflow of cell dissociation, FACS isolation of Ctnnb1Het (Control) and ∆Ctnnb1 Cxcl12-tdT+ cells and bulk RNA-seq. b FACS sorting of Cxcl12-creER+ AP cells at P10 (pulsed at P3). Blue box: Cxcl12CE-Ctnnb1Het-P3 (Ctnnb1Het) cells isolated from AP-Ctnnb1 cHet molars, red box: Cxcl12CE-∆Ctnnb1-P3 (∆Ctnnb1) cells isolated from AP-Ctnnb1 cKO molars. c–f RNA-seq analysis results. c Heatmaps of representative 20 differentially expressed genes (DEGs) associated with non-canonical Wnt signaling, Wnt antagonist, chondrocyte, periodontal ligament (PDL), osteoblast, and dental pulp/odontoblast (DP/Od) markers, as well as chemokine. Left 4 lanes: Ctnnb1Het, right 4 lanes: ∆Ctnnb1. PDL: periodontal ligament, DP: dental pulp, Od: odontoblast. Yellow: higher expression, purple: lower expression, n = 4 mice per each group. d Top enriched GO terms – biological processes (GO-BP) overrepresented in DEGs (Adjusted p < 0.01, Fisher’s exact test). e, f GSEA analysis. Representative 4 pathways (e) and 2 up-regulated signaling pathways (f) in Cxcl12CE-∆Ctnnb1-P3 cells. NES: normalized enrichment score (Adjusted p < 0.001, weighted Kolmogorov-Smirnov test). g, h Administration of TGF-β receptor inhibitor galunisertib (Gal) to AP-Ctnnb1 cHet and cKO mice. Gal or PBS was administrated into AP-Ctnnb1 cHet or cKO from P8 to P20. g Mandibular 1st molar of AP-Ctnnb1 cHet and cKO at P25 (pulsed at P3). Scale bars: 500 μm (upper panels), 100 μm (lower panels). h Quantification of POSTN+tdTomato+ cells/tdTomato+ dental pulp cells. n = 3 mice (AP-Ctnnb cHet + PBS), n = 4 mice (AP-Ctnnb cHet + Gal), n = 6 mice (AP-Ctnnb cKO + PBS), n = 4 mice (AP-Ctnnb cKO + Gal), *P < 0.05, **P < 0.01, ***P < 0.001, Two-tailed, one-way ANOVA followed by the Mann-Whitney U test. All data are mean ± SD. Exact P-value is indicated in the figures. Source data are provided as a Source Data file.
