Fig. 3

Effects of SATB2 on osteo/odontogenic differentiation of hDPSCs. a Calcium nodule deposition of hDPSCs examined by Alizarin Red S staining showed that SATB2 knockdown significantly decreased the capacity of osteo/odontogenic differentiation of hDPSCs compared with the control group. b Quantitative RT-PCR revealed that the mRNA level of RUNX2, OPN, SEMA7A, SP7, DLX and ALP, but not IGFBP3, decreased after knockdown of SATB2. c Western blot analysis showed that RUNX2 was significantly downregulated after SATB2 knockdown compared with the control ones. d The DPSCs transfected with wild-type SATB2 showed increased capacity of osteo/odontogenic differentiation compared to vehicle, while cells transfected with mutant SATB2 showed decreased capacity of osteo/odontogenic differentiation compared to the ones transfected with wild-type SATB2. e Transfection of wild-type SATB2 upregulated the expression of RUNX2, OPN, SEMA7A, SP7, DLX and ALP, but not IGFBP3, compared with vehicle. Cells transfected with mutant SATB2 expressed lower level of RUNX2, OPN, SP7 and DLX3 compared with wild-type. f Western blot analysis showed that RUNX2 was up-regulated when hDPSCs were transfected with wild-type SATB2, while mutant SATB2 transfection showed lower expression level of RUNX2 compared with the wild-type. Each experiment was repeated three times with n ≥ 3 samples per group. Scale bar 200 μm, *P < 0.05, **P < 0.01, ***P < 0.001. Data are expressed as the means + SD. si NC: negative control siRNA transfection, si SATB2: SATB2 siRNA transfection, Vehicle: vector transfection, WT SATB2: wild-type SATB2 transfection, MU SATB2: mutant SATB2 transfection