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Fig. 8 | Stem Cell Research & Therapy

Fig. 8

From: Effects of smoking on the tissue regeneration-associated functions of human endometrial stem cells via a novel target gene SERPINB2

Fig. 8

Cigarette smoke exposure markedly suppressed various tissue regeneration-associated functions of endometrial stem cells in vivo. Schematic of the in vivo experimental procedure used, which is described in detail in Materials and Methods (A). Mice were intraperitoneally administrated cigarette smoke extract 10 times at 0.5 or 1 mg/kg mg/kg for or vehicle. Mice endometrial stem cells were isolated from uterine tissues, and the inhibitory effects of cigarette smoke on cell viability were assessed using an MTT assay (B). The changes in stem cell migratory abilities were analyzed using a transwell assay (C) and by western blotting using MMP-2 and MMP-9 antibodies (D). The inhibitory effects of cigarette smoke exposure on adipocyte (E) and osteoblast (F) differentiation of mouse endometrial stem cells in vivo were analyzed by oil red O and alizarin red staining, respectively. Relative quantifications of calcium mineral content and lipid droplet formation within differentiated cells were evaluated by measuring absorbance at 500 and 570 nm, respectively. The inhibitory effects of cigarette smoke exposure on the expression of various pluripotency-associated genes, C-MYC, KLF4, NANOG, OCT4, and SOX2 were analyzed by real-time PCR (G). β-actin was used as the internal control, and PPIA as the housekeeping gene for real-time PCR. 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)

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