1. Interleukin-1β induces cyclooxygenase-2 expression and promotes the invasive ability of human mesenchymal stem cells derived from ovarian endometrioma 
An-Pei Kao, M.S., Kai-Hung Wang, Ph.D., Cheng-Yu Long, M.D., Ph.D., Chee-Yin Chai, M.D., Ph.D., Cheng-Fang Tsai, B.S., Tsung-Hua Hsieh, M.S., Chia-Yi Hsu, M.S., Chia-Cheng Chang, Ph.D., Jau-Nan Lee, M.D., Ph.D., Eing-Mei Tsai, M.D., Ph.D. 
Fertility and Sterility 
Volume 96, Issue 3, Pages e1 (September 2011)
DOI: /j.fertnstert
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
(A) The messenger RNA (mRNA) expressions of eutopic and ectopic endometrial mesenchymal stem cells (EN-MSCs) compared in a scatter plot. They presented similar patterns in Pearson correlation R2 of (B) Two identified genes, interleukin-1β (IL-1β) and IL-8, which simultaneously related to metastasis, angiogenesis, and inflammation. The expressions of IL-1β and cyclooxygenase-2 (COX-2) genes were statistically significantly higher in ectopic EN-MSCs than in eutopic EN-MSCs, as analyzed by (C) reverse-transcription polymerase chain reaction (RT-PCR) and (D) quantitative qPCR (∗P<.05).
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
(A) Although both eutopic and ectopic endometrial mesenchymal stem cells (EN-MSCs) expressed cyclooxygenase-2 (COX-2) protein by Western blot analysis, the expression level was higher in the latter. The COX-2 protein expression was blocked by treatment with U0126 (10 μM), a highly selective inhibitor of MAPK/ERK kinase, in a time-dependent manner and NS398 (100 μM), a COX-2 inhibitor, for 24 hours. Ectopic EN-MSCs showed statistically significantly higher (B) migration and (C) invasion ability than eutopic EN-MSCs. Both abilities of ectopic EN-MSCs could be statistically significantly blocked by U0126 and NS398 treatments (∗P<.05). The significant inhibiting effects on migration and invasion by these two inhibitors were also observed for eutopic EN-MSCs (not indicated in figures).
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
(A) After treatment with interleukin-1β (IL-1β, 1 ng/mL) for 24 hours, the expression of COX-2 protein was enhanced in both eutopic and ectopic endometrial mesenchymal stem cells (EN-MSCs) in a time-dependent manner, as shown by Western blot analysis. The increase was more dramatic in the latter. (B) Ectopic EN-MSCs showed statistically significantly higher migration ability than eutopic EN-MSCs. (see Fig. 2B). This cell migration ability of ectopic EN-MSCs can be further enhanced by IL-1β treatment (1 ng/mL, 24 hours) and can be statistically significantly inhibited by treatment with U0126 (10 μM) or NS398 (100 μM) for 24 hours (∗P<.05; both inhibitor treatments compared with no treatment control or IL-1β–treated cells). (C) The invasion ability of ectopic EN-MSCs was statistically significantly higher than that of eutopic EN-MSCs (see Fig. 2C). The invasion ability of ectopic EN-MSCs can be further enhanced by IL-1β treatment and suppressed by U0126 or NS398 treatments (∗P<.05; both inhibitor treatments compared with no treatment control or IL-1β–treated cells). (D) A diagram shows the development of ectopic EN-MSC organoids on Matrigel-coated plates in the presence of IL-1β. These ectopic EN-MSCs were prepared at high concentration (1 × 107 cells/mL), and 5 × 104 cells in 5 μL medium were inoculated in a Matrigel-coated plate. After plating for 2 hours, these cell masses were covered with culture medium and incubated. (E) After plating for 45 days, the organoid spheres of group 1 (control without IL-1β) did not change much in size. After culture for 60 days, the spheres showed a slightly rugged surface. In contrast, the organoids of group 2 (with IL-1β treatment) became irregular spheres in 45 days. After incubation for 60 days, these organoids were statistically significantly larger than those in group 1 and were found to develop many tentacle-like arms that extended into the Matrigel.
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

5. Supplemental Figure 1
(A) Compared with eutopic endometrial mesenchymal stem cells (EN-MSCs), ectopic EN-MSCs showed a high level of gene expression in immune response pathways in the top 10 GeneGo pathway maps by MetaCore analysis. (B) Three inflammation and two cell-adhesion–related processes were highly expressed in ectopic EN-MSCs among the top 10 GeneGo process networks.
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions