1. Leiomyoma-derived transforming growth factor-β impairs bone morphogenetic protein-2- mediated endometrial receptivity 
Leo F. Doherty, M.D., Hugh S. Taylor, M.D. 
Fertility and Sterility 
Volume 103, Issue 3, Pages (March 2015)
DOI: /j.fertnstert
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
The ELISA quantification of transforming growth factor (TGF)-β3 in endometrial stromal cell (ESC)-conditioned and leiomyoma-conditioned media. The TGF-β3 protein concentration was quantified by ELISA in conditioned media from primary ESC and leiomyoma cells grown at 60%–70% confluence and cultured for 24 hours. Mean TGF-β3 protein concentration was significantly higher in leiomyoma-conditioned media compared with ESC-conditioned media (52 vs. 9.2 ng/mL; ∗P<.05). The TGF-β3 protein was present in low concentration (3.2 ng/mL) in standard media (Dulbecco's minimum essential medium [DMEM] containing 1% penicillin, 1% streptomycin, 1% amphotericin B, and 10% fetal bovine serum).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Expression of bone morphogenetic protein (BMP) receptors in leiomyoma-conditioned media (LCM) exposed endometrial stromal cells (ESCs). (A) The BMP receptor 2. BMPR2 expression was reduced in ESC exposed to LCM compared with control non-LCM-exposed ESC (0.59-fold; P<.05). Preincubation of LCM with transforming growth factor-β neutralizing antibody, before ESC exposure, prevented repression of BMPR2 expression. (B) The BMP receptor 1B. BMPR1B expression was reduced in ESC exposed to LCM compared with control non-LCM-exposed ESC (0.61-fold; P<.05). Preincubation of LCM with transforming growth factor-β neutralizing antibody, before ESC exposure, prevented repression of BMPR1B expression. (C) The BMP receptor 1A. BMPR1A expression was not significantly changed in LCM-exposed ESC compared with control non-LCM-exposed ESC. ∗denotes statistical significance with P<.05.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Bone morphogenetic protein (BMP)-2 stimulated HOXA10 and LIF expression in leiomyoma-conditioned media (LCM)-treated endometrial stromal cells (ESCs). Effect of transforming growth factor (TGF)-β antibody. The BMP-2 responsiveness was assessed in LCM-exposed ESC by treatment with recombinant human BMP-2. HOXA10 and leukemia inhibitory factor (LIF) expression were quantified by quantitative real-time polymerase chain reaction (PCR) after 24 hours of recombinant human BMP-2 treatment. (A) The BMP-2-stimulated HOXA10 expression. The BMP-2-stimulated HOXA10 expression was unchanged in myometrium-conditioned media (MCM) exposed ESC compared with nonexposed control ESC (1.07-fold; P=not significant [NS]). The BMP-2-stimulated HOXA10 expression was repressed in ESC exposed to LCM compared with non-LCM-exposed control ESC (0.63-fold; P<.05). Incubation of LCM with TGF-β neutralizing antibody, before ESC exposure, prevented repression of BMP-2-stimulated HOXA10 expression. (B) The BMP-2-stimulated LIF expression. The BMP-2-stimulated LIF expression was unchanged in MCM-exposed ESC compared with nonexposed control ESC (0.94-fold; P=NS). The BMP-2-stimulated LIF expression was repressed in LCM-exposed ESC compared with non-LCM-exposed control ESC (0.29-fold; P<.05). Incubation of LCM with TGF-β neutralizing antibody, before ESC exposure, prevented repression of BMP-2-stimulated LIF expression. ∗denotes statistical significance with P<.05.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Bone morphogenetic protein (BMP)-2-stimulated HOXA10 and leukemia inhibitory factor (LIF) expression in leiomyoma-conditioned media (LCM)-treated endometrial stromal cells (ESCs). Effect of transfection with mutant transforming growth factor (TGF)-β receptor. The BMP-2-stimulated expression of HOXA10 and LIF was examined in LCM exposed ESCs that had been transfected with mutant TGF-β type II receptor or vector control. (A) The BMP-2-stimulated HOXA10 expression. The BMP-2-stimulated HOXA10 expression was repressed in LCM-exposed ESC that had been transfected with vector control plasmid compared with non-LCM-exposed controls (0.33-fold; P<.05). The BMP-2-stimulated HOXA10 expression was not significantly changed compared with non-LCM-exposed control ESCs when ESCs had been transfected with mutant TGF-β type II receptor before LCM exposure. (B) The BMP-2-stimulated LIF expression. The BMP-2-stimulated LIF expression was repressed in LCM-exposed ESCs that had been transfected with vector control plasmid compared with non-LCM-exposed control ESCs (0.64-fold; P<.05). The BMP-2-stimulated LIF expression was significantly increased compared with non-LCM-exposed control ESCs when ESCs had been transfected with mutant TGF-β type II receptor before LCM exposure (1.57-fold; P<.05). ∗denotes statistical significance with P<.05.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions