Fig. 5 | International Journal of Oral Science

Fig. 5

From: Atp6i deficient mouse model uncovers transforming growth factor-β1 /Smad2/3 as a key signaling pathway regulating odontoblast differentiation and tooth root formation

Fig. 5The alternative text for this image may have been generated using AI.

Atp6i deficient osteoclastic bone resorption-conditioned medium inhibits odontoblast differentiation and TGF-β1/ Smad2/3 signaling transduction in vitro. a, b Immunofluorescent staining of DSP and OSX in A4 pulp precursor cell line induced by different cell culture media for 3 days. c qPCR analysis of gene expression levels of odontoblast differentiation markers in odontoblast-like cells induced by different cell culture media for 7 days. Hprt was used as an endogenous control. d Immunofluorescent staining of pSmad2/3 in odontoblast-like cells induced by different cell culture media for 3 days. e Western blot of p-Smad2/3 expression in odontoblast-like cells induced by different cell culture media for 7 days. f Active TGF-β1 level in different cell culture media detected by Elisa; WT OC + bone CM versus other groups. g qPCR analysis of odontoblast differentiation markers gene expression levels in odontoblast-like cells induced by WT OC BRCM with addition of TGF-β1 neutralization antibody for 7 days. WT OC BRCM added with IgG was used as the control group. WT OC CM, WT osteoclast culture medium; WT OC + bone CM (WT BRCM), conditioned medium from WT osteoclast cultured with bone; Atp6i−/− OC + bone CM, conditioned medium from Atp6i−/− osteoclast cultured with bone; WT OC + bone CM with TGF-β1 Ab, WT OC BRCM added with TGF-β1 neutralization antibody. n = 7 in each group; *P < 0.05; **P < 0.01; ns, not significant. h Heatmap showing expression levels of odontoblast markers in P20 WT and Atp6i−/− mice mandibles

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