Fig. 6

Effects of SERPINB2 overexpression on various tissue regeneration-associated functions of endometrial stem cells. Schematic showing the regulatory roles of SERPINB2 on various tissue regeneration-associated functions of endometrial stem cells (A). Transfection of endometrial stem cells with a specific retroviral expression vector for SERPINB2 significantly inhibition inhibited cell proliferation as compared with control vector transfection (B). Increased levels of caspase-3 activities following SERPINB2 overexpression were analyzed by western blotting (C), and SERPINB2 overexpression-induced apoptotic DNA fragmentations in endometrial stem cells were also analyzed by DAPI staining (D). The inhibitory effects of SERPINB2 overexpression on the migration capacity of endometrial stem cells were assessed by transwell assay (E) and western blotting using specific MMP-2 and MMP-9 antibodies (F). The inhibitory effects of SERPINB2 overexpression on osteogenic (G) and adipogenic (H) differentiation were analyzed by alizarin red or oil red O staining, respectively. Relative quantifications of calcium mineral contents and lipid droplet formation were determined by measuring absorbance at 570 or 500 nm, respectively. The inhibitory effects of SERPINB2 overexpression on the expressions of pluripotency-associated genes C-MYC, KLF4, NANOG, OCT4, and SOX2 were analyzed by real-time PCR (I). β-actin was used as an internal control, and PPIA as the housekeeping gene for real-time PCR analysis. All experiments were performed in triplicate. Data are presented as means ± SDs. *, p < 0.05; **, p < 0.005; and ***, p < 0.001 (two-sample t test)