Fig. 5

SERPINB2 mediated the harmful effects of cigarette smoke exposure on various tissue regeneration-associated functions of endometrial stem cells. Schematic diagram describing the roles of SERPINB2 as a key regulator that mediates cigarette smoke-induced harmful effects in endometrial stem cells (A). Cells were transfected with shRNA specifically targeting SERPINB2 and then exposed or not to 1% cigarette smoke extract for 72 h. Changes in cell viability were determined using an MTT assay (B). SERPINB2 knockdown attenuated cigarette smoke-mediated inhibition of endometrial stem cells migration as determined by transwell assay (C) and western blotting using MMP-2 and MMP-9 antibodies (D). Cells were transfected with a SERPINB2 shRNA and then treated with or without 1% cigarette smoke extract; subsequent changes in adipocyte (E) and osteoblast (F) differentiation abilities were analyzed by oil red O and alizarin red staining, respectively. Relative calcium mineral contents and lipid droplet formation within differentiated cells were assessed by measuring absorbance at 500 and 570 nm, respectively. Blocking of the effects of SERPINB2 knockdown on the cigarette smoke-induced inhibitions of pluripotency-associated factors C-MYC, KLF4, NANOG, OCT4, and SOX2 was investigated by real-time PCR (G). β-actin was used as the 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)