Fig. 7: USP49 localization and its effects on the expression profiles of Pax9 and Msx1 during different stages of tooth development.

A USP49 expression patterns during tooth development at the bud, cap, and bell stages. USP49 was expressed in the epithelium and mesenchyme (a) at the bud stage and in the developing bone surrounding the developing tooth germs at both the (b) cap and (c) bell stages. (d) Quantification of the fluorescence intensity of USP49 in developing dental epithelium and dental mesenchyme of tooth germs. Scale bar: 100 µm. Six biological replicates at each stage (n = 6) (fluorescence intensities were quantified from three biological replicates with randomly selected sections at each stage). The data presented here represent the mean ± SD of three independent experiments (**P < 0.001 and ***P < 0.0001). Statistical significance was analyzed by ANOVA followed by Tukey’s post hoc test. B Expression levels of Msx1 and Pax9 in tooth germs (a, d) mock-group and (b, e) sgRNA1-USP49-depletion group after 2 days of in vitro culture. The fluorescence intensities of (c) Msx1 and (f) Pax9 were compared between the mock-group and USP49-depletion group (n = 3). Scale bar = 100 µm. Epi = Epithelium, Mes = Mesenchyme, DB = Developing Bone. Six biological replicates per group (n = 6) (fluorescence intensities were quantified from three biological replicates with randomly selected sections at each stage). The data presented here represent the mean ± SD of three independent experiments (***P < 0.0001) by Student’s t test.