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. 2022 Jun 21;10:890882. doi: 10.3389/fbioe.2022.890882

FIGURE 5.

FIGURE 5

Role of Epfn activation at stages 2 and 3 in ameloblast induction. (A) Diagram of ameloblast induction from Epfn-iPSCs. Dox (1 μg/ml) was added at stages 2 and 3 to activate Epfn expression. (B) Cell morphology (upper) and mCherry expression (lower) on day 17. Scale bar: 100 μm. (C) WST-1 test on day 17. **, p < 0.01. (D) ARS stain on day 17. Scale bar: 1 cm. (E) Quantification of ARS staining on day 17. **, p < 0.01 (t-test; n = 3). (F) Gene expression according to RT-PCR analysis on day 17. Markers (stem cells: Oct4; dental epithelial: Krt14, p75, and tuftelin; and ameloblast: Amelx, Ambn and KLK4). (G) Relative quantitative analysis of RT-PCR results. **, p < 0.01 (t-test; n = 3). (H) Western blotting on day 17. (I) Relative quantitative analysis of Western blot results. *, p < 0.05; **, p < 0.01 (t-test; n = 3). (J) Real-time PCR of the BMP-Smad signaling pathway-associated molecules on day 17. **, p < 0.01 (t-test; n = 3).