Hypoxia inducible factor and microvessels in peri-implantation endometrium of women with recurrent miscarriage Xiaoyan Chen, M.Phil., Lingming Jiang,

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Hypoxia inducible factor and microvessels in peri-implantation endometrium of women with recurrent miscarriage Xiaoyan Chen, M.Phil., Lingming Jiang, M.D., Chi Chiu Wang, Ph.D., Jin Huang, Ph.D., Tin Chiu Li, M.D., Ph.D. Fertility and Sterility Volume 105, Issue 6, Pages 1496-1502.e4 (June 2016) DOI: 10.1016/j.fertnstert.2016.02.032 Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Hypoxia inducible factor 1α (HIF1α) expression in the endometrium from (A) women with recurrent miscarriage (n = 24) and (B) normal fertile women (n = 36). Original magnification, ×200. Scale bar = 100 μm. GE = glandular epithelium; LE = luminal epithelium; SC = stromal cell. Fertility and Sterility 2016 105, 1496-1502.e4DOI: (10.1016/j.fertnstert.2016.02.032) Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions

Figure 2 The micro–blood vessels in endometrial biopsies were determined by anti-von Willebrand factor (vWF)–stained sections. Representative images of blood vessels localized in endometrium (arrows) from (A) women with recurrent miscarriage (n = 24) and (B) healthy fertile women (n = 36) are shown. Original magnification, ×200. Scale bar = 100 μm. Fertility and Sterility 2016 105, 1496-1502.e4DOI: (10.1016/j.fertnstert.2016.02.032) Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Women with recurrent miscarriage had an increased (A) number and (C) volume of micro–blood vessels compared with healthy fertile women. (B) There was no statistically significant difference in the mean diameter of blood vessels between two groups. **P<.05. Fertility and Sterility 2016 105, 1496-1502.e4DOI: (10.1016/j.fertnstert.2016.02.032) Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Specificity of anti–hypoxia inducible factor 1α (HIF1α) antibody. (A) Representative Western blot of HIF1α in human endometrium lysate from normal individuals (n = 3). Anti-HIF1α antibody recognizes a single band representing HIF1α. (B) Western blot of HIF1α in endometrium lysate with recombinant HIF1α preabsorbed antibody. Preincubation with recombinant HIF1α decreased HIF1α signal. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Fertility and Sterility 2016 105, 1496-1502.e4DOI: (10.1016/j.fertnstert.2016.02.032) Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 2 Specificity of anti-von Willebrand factor (vWF) antibody. (A) Representative Western blot of vWF in human endometrium lysate from normal individuals (n = 3). Anti-vWF antibody recognizes a single band representing vWF. (B) Western blot of vWF in endometrium lysate with recombinant vWF preabsorbed antibody. Preincubation with recombinant vWF decreased the vWF signal. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Fertility and Sterility 2016 105, 1496-1502.e4DOI: (10.1016/j.fertnstert.2016.02.032) Copyright © 2016 American Society for Reproductive Medicine Terms and Conditions