Fig 2: The depletion of KMT5B enhanced the osteo/dentinogenic differentiation of DPSCs under normoxia and hypoxia. a The knockdown efficiency of KMT5B by western blot detection. b ALP activity assay. c Alizarin Red S staining. d Quantitative calcium measurements. e The OCN, DSPP, DMP1, and BSP expressions at 7 days after the induction in DPSCs by western blot detection. f–i 50 μmol·L−1 CoCl2 was used to treat DPSCs. f ALP activity assay. g Alizarin Red S staining. h Quantitative calcium measurements. i The OCN, DSPP, DMP1, and BSP expressions at 7 days after the induction in DPSCs by western blot detection. The one-way ANOVA or Student’s t-test was applied to confirming statistical significance. GAPDH was acted as an internal control. Error Bars mean standard deviations (n = 3). **P ≤ 0.01

Fig 3: Knockdown of miR-140-3p repressed the osteo/dentinogenic differentiation of DPSCs. a Real-time RT-PCR exhibited the knockdown efficiency of miR-140-3p in DPSCs. b ALP activity assay. c Alizarin Red S staining. d Quantitative calcium measurements. e The OCN, DSPP, DMP1, and BSP expressions at 7 days after the induction in DPSCs. Student’s t-test was applied to confirming statistical significance. Histone H3 or U6 was acted as an internal control. Error Bars mean standard deviations (n = 3). *P ≤ 0.05; **P ≤ 0.01

Fig 4: Overexpression of KMT5B repressed the osteo/dentinogenic differentiation of DPSCs. a Western blot exhibited the overexpression of KMT5B in DPSCs. b ALP activity assay. c Alizarin Red S staining. d Quantitative calcium measurements. e The OCN, DSPP, DMP1, and BSP expressions at 7 days after the induction in DPSCs by western blot detection. Student’s t-test was applied to confirming statistical significance. GAPDH was acted as an internal control. Error Bars mean standard deviations (n = 3). *P ≤ 0.05; **P ≤ 0.01

Fig 5: Imunostain validations of RNA-Seq predicted gene expression patterns.The tissues sections shown are of one half of the E14.5 forming palate, with the medial edge epithelium at the left side of the image. For Agtr2 the RNA-Seq strongest expression was in the lateral and oral compartments, consistent with the immunostain results. Crabp1 immunostain showed strongest expression in the lateral compartment, as predicted by RNA-Seq. Both App and Ibsp showed strongest expression in the nasal compartment (above), compared to oral compartment (below), as predicted by RNA-Seq.