1. Peroxisome proliferator-activated receptor-γ ligand reduced tumor necrosis factor-α- induced interleukin-8 production and growth in endometriotic stromal cells 
Yoko Ohama, M.D., Tasuku Harada, M.D., Tomio Iwabe, M.D., Fuminori Taniguchi, M.D., Yasuko Takenaka, M.D., Naoki Terakawa, M.D. 
Fertility and Sterility 
Volume 89, Issue 2, Pages (February 2008)
DOI: /j.fertnstert
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Expression of peroxisome proliferator-activated receptor-γ (PPARγ) in endometriotic stromal cells. (A) The polymerase chain reaction revealed PPARγ (479 bp) mRNA expression. Endometrial stromal cells were used as positive controls and glycerol-3-phosphate dehydrogenase (G3PDH) (450 bp) was used as an internal control for RNA loading. RT (−) was the absence of reverse transcriptase. (B1) Immunocytochemical staining revealed PPARγ protein in endometrial stromal cells (positive control). (B2) Endometriotic stromal cells. PPARγ protein was predominantly located in the cell nucleus. (B3) Negative control (with elimination of the primary antibody). Original magnification, ×400.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Effect of pioglitazone on interleukin-8 protein production in endometriotic stromal cells. Adding tumor necrosis factor-α (TNF-α; 100 pg/mL) for 24 hours significantly increased interleukin-8 protein. Results were expressed as a percentage of control values. Treatment of endometriotic stromal cells with 0.1–10 μM of pioglitazone significantly reduced the TNF-α-induced interleukin-8 production. Concentrations of interleukin-8 protein in supernatants were measured by ELISA. The minimum detectable level was 3.5 pg/mL. Bars represent SE. The means and SE were calculated from different patient's endometriotic stromal cell cultures (n = 8). The cells were used for experiments during one- to two-cell passages. ∗P<.01 versus adding TNF-α.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Effect of pioglitazone on endometriotic stromal cell proliferation. Cell proliferation was determined by the 5-bromo-2′-deoxyuridine proliferation assay. The number of endometriotic stromal cells increased in the presence of tumor necrosis factor-α (TNF-α; 100 pg/mL), and this bar shows as a control. Results were expressed as a percentage of control values. Adding 0.1–10 μM of pioglitazone significantly suppressed the growth of endometriotic stromal cells. Bars represent SE. The means and SE were calculated from different patient's endometriotic stromal cell cultures (n = 5). The cells were used for experiments during one- to two-cell passages. ∗P<.01 versus adding TNF-α.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Effect of pioglitazone on expression of p-IκB protein and the concentration of intranuclear p65 protein in endometriotic stromal cells. (A) The confluent endometriotic stromal cells were exposed to media with tumor necrosis factor-α (TNF-α) (100 pg/mL) and pioglitazone (10 μM) for 10 minutes. Proteins were separated by electrophoresis on 4%–20% sodium dodecyl sulfate gels and immunoblotted using anti p-IκBα (Ser32) antibody. TNF-α increased the expression of phosphorylation of IκB. Adding pioglitazone (10 μM) did not influence the expression of p-IκB (A1). Relative densitometric units of the band are shown in the lower panel, with the density of the control band (without both TNF-α and pioglitazone) set arbitrarily at 1.0. The data are the means with SE of three different experiments (A1, A2). (B) The concentration of intranuclear p65 protein was determined in duplicate using nuclear factor (NF)-κB transcription factor assay ELISA. TNF-α (100 pg/mL) markedly increased the intranuclear concentration of p65. Adding pioglitazone significantly reduced the concentration of p65.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions