1. Expression of the pluripotent transcription factor OCT4 promotes cell migration in endometriosis 
Jui-Hung Chang, Ph.D., Heng-Kien Au, M.D., Wei-Chin Lee, M.S., Ching-Chi Chi, M.D., Ph.D., Thai-Yen Ling, Ph.D., Le-Ming Wang, M.D., Shu-Huei Kao, Ph.D., Yen-Hua Huang, Ph.D., Chii-Ruey Tzeng, M.D. 
Fertility and Sterility 
Volume 99, Issue 5, Pages e5 (April 2013)
DOI: /j.fertnstert
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Quantification of OCT4 messenger RNA expression in human endometrial tissues. Reverse transcription and quantitative real-time polymerase chain reaction (PCR) were used to quantify the levels of (A) OCT4 and (B) NANOG messenger RNAs in primary endometriotic stromal cells, normal endometrium (n = 9), normal myometrium (n = 3), hyperplastic endometrium (n = 36), adenomyosis (n = 30), and chocolate cysts (n = 28). The RL95-2 cells were used as an internal control for normalization. Results are expressed as the mean ± SD. P<.0001 compared with the hyperplasia group.
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3. Figure 2
Positive correlations of the expression of OCT4 and that of migration-associated genes in human endometriotic tissues. The (A) VIMENTIN, (B) TWIST, (C) SNAIL, and (D) SLUG messenger RNA levels in endometriotic tissue samples from adenomyosis (n = 30) and chocolate cysts (n = 28) were quantified using reverse transcription and quantitative real-time polymerase chain reaction (PCR). The transcriptional levels (ΔCt of gene level/β2-microglobulin) of the migration-associated genes were compared to that of OCT4 using the Pearson correlation analysis to determine statistical correlations. (A) adenomyosis, P=.447; chocolate cysts, P<.01; (B) adenomyosis, P<.001; chocolate cysts, P<.0001; (C) adenomyosis, P<.005; chocolate cysts, P<.0001; (D) adenomyosis, P<.0001; chocolate cysts, P<.0001.
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Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
The OCT4 increases expression levels of migration-associated genes and migratory ability of human endometrial cells. (A) The relative transcriptional levels of VIMENTIN, TWIST, SNAIL, SLUG, and N-CADHERIN (N-CAD) in RL95-2, HEC1A, and primary endometriotic stroma cells with or without OCT4 overexpression were determined using reverse transcription and quantitative real-time polymerase chain reaction (PCR). (B) The relative levels of migration-associated proteins in RL95-2 and HEC1A cells with or without OCT4 overexpression were analyzed by Western blotting. Data are representative of at least three independent experiments. (C) The effect of OCT4 expression on the motility of human endometrium cells at 0 and 48 hours was quantified using a transwell assay. (D) The effect of OCT4 expression on cell migration of human endometrial cells at 0, 24, and 48 hours was evaluated using a wound closure assay. The percentage of the area in the wound closure assay with or without OCT4 overexpression is shown. (E) The cellular localization of actin protein in human endometrial RL-95-2 and HEC1A cell lines and primary stromal cells with or without OCT4 protein overexpression was evaluated using immunocytochemical staining. Nuclei were stained with 6-diamino-2-phenylindole (blue), and actin was detected with fluorescein isothiocyanate conjugate (green). Higher magnification and phase-contrast images are shown. *P<.05 by t test.
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5. Supplemental Figure 1
Relative protein expression levels of migration-associated proteins in RL95-2 and HEC1A cells with or without OCT4 overexpression. Western blot analysis (Fig. 3B) was repeated at least three times independently. The relative levels of protein in OCT4 overexpressing cells are normalized to the control group. *P<.05 by t test.
Fertility and Sterility  , e5DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

6. Supplemental Figure 2
Effects of OCT4 overexpression on cell proliferation in human endometrial RL95-2 and HEC1A cell lines and primary endometriotic stromal cells. The viability of human endometriotic cells with or without OCT4 overexpression was evaluated using a WST-1 assay. Three independent experiments were performed for each experimental condition.
Fertility and Sterility  , e5DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions