1. Changes of human decidual natural killer cells cocultured with YFP-Toxoplasma gondii: implications for abnormal pregnancy 
Xiaoyan Xu, M.D., Qiang Fu, M.D., Qun Zhang, M.D., Mingdong Zhao, M.D., Zonghua Gao, M.D., Xianbing Liu, M.D., Yang Liu, M.D., Xuemei Hu, M.D., Ph.D. 
Fertility and Sterility 
Volume 99, Issue 2, Pages e2 (February 2013)
DOI: /j.fertnstert
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Expression of KIR2DL4, ILT-2, and NKG2D on decidual natural killer cells is altered by infection with Toxoplasma gondii. After 12, 24, and 48 hours of infection, the expression levels of (A) KIR2DL4, (B) ILT-2, and (C) NKG2D on the surface of uninfected and infected decidual natural killer cells were analyzed by flow cytometry. Data are presented as the means ± SEM, n = 30, P<.01.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
The cytotoxicity caused by Toxoplasma gondii was measured by costaining uninfected and infected decidual natural killer (dNK) cells with PKH67 and To-Pro-3 followed by flow cytometry. (A) Total human extravillous trophoblast cells were stained with PKH67. Lysed cells are observed in the upper right frame. (B) Percentages of lysed human extravillous trophoblast cells in the presence of dNK cells incubated for 12, 24, or 48 hours with T. gondii. Cytotoxicity of uninfected dNK cells was also determined. Data shown are averages of three independent experiments.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

4. Supplemental Figure 1
Purified decidual natural killer (dNK) cells were identified and phenotypic analysis of isolated CD3−CD56+ human dNK cells using flow cytometry. The dNK cells were isolated from fresh human decidual tissues and were divided into two parts: one part of dNK cell was stained with anti-CD56-PE and anti-CD3-FITC antibodies and the other part of dNK cells was stained using anti-CD56-PE and anti-CD16-PE-cy5 antibodies, and then analyzed by flow cytometry. (A) More than 95% cells were CD3−CD56+ dNK cells. (B) The upper left frame shows the predominant CD56brightCD16− dNK subset, and the upper right frame shows the minor CD56dimCD16+ dNK subset. The percentages of their populations in the total isolated cells are indicated.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

5. Supplement Figure 2
Changes of subset of decidual natural killer (dNK) cells after cocultured with Toxoplasma gondii. (A) The morphology of uninfected dNK cells as observed by fluorescence microscopy (×400). (B) The morphology of dNK cells after 48 hours in culture with YFP-T. gondii as observed by fluorescence microscope. The infection rate was about 75%. (C) The infected dNK cells, stained with anti-CD56-PE antibody, were detected by confocal microscopy. The YFP-T. gondii infected cells were marked with arrows. (D) Uninfected dNK cells and (E) infected dNK cells were stained with anti-human CD16-PE-cy5 antibody and analyzed by flow cytometry.
Fertility and Sterility  , e2DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions