Fig. 2

Identification of human dental pulp stem cells and influence of SATB2 on proliferation rate of hDPSCs. a Flow cytometry assays showed that hDPSCs were positive for CD29 and CD73, and negative for CD34 and CD45. b Cell immunofluorescence showed that intrinsic SATB2 expressed exclusively in the nucleus. c Western blot analysis showed that the molecular weight of mutant SATB2 was significantly lower than the wild-type. d Cell immunofluorescence of transfected flag-tagged SATB2 in hDPSCs. Wild-type SATB2 located in the nucleus while mutant SATB2 was distributed over the cytoplasm. Transfection efficiency of both wild-type and mutant SATB2 was around 80%. e BrdU-labeling assay showed that the proliferation rate of hDPSCs transfected with SATB2 siRNA was lower than negative control. f BrdU-labeling assay showed that the proliferation rate of hDPSCs transfected with mutant SATB2 was lower than cells transfected with wild-type. g SATB2 knockdown in hDPSCs resulted in decreased expression of ATF4, BSP and COL1A1. Transfection of wild-type SATB2 upregulated the expression of ATF4, BSP and COL1A1 compared with vehicle, while cells transfected with mutant SATB2 expressed lower level of ATF4, BSP and COL1A1 compared with wild-type. Data are expressed as the means + SD. Each experiment was repeated three times with n ≥ 3 samples per group. Scale bar 20 μm, *P < 0.05, ** P < 0.01, *** P < 0.001, NS non-significant. 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