Haptoglobin β chain isoforms in the plasma and peritoneal fluid of women with endometriosis  Simone Ferrero, M.D., David John Gillott, Ph.D., Valentino Remorgida, M.D., Paola Anserini, M.D., Karen Price, Ph.D., Nicola Ragni, M.D., Jurgis Gediminas Grudzinskas, M.D.  Fertility and Sterility  Volume 83, Issue 5, Pages 1536-1543 (May 2005) DOI: 10.1016/j.fertnstert.2004.11.064 Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Volume of the Hp β chain isoforms in relation to the amount of proteins (range, 30–180 μg) loaded in the two-dimensional gels. The gels were silver stained, and the Vo was calculated by using the Phoretix 2D v5.01 software. The Vo of the Hp β chain isoforms was directly proportional to the protein quantity loaded on the gels with all ρ greater than 0.802; the HpβE spot had ρ = 0.961 (Pearson's correlation). Fertility and Sterility 2005 83, 1536-1543DOI: (10.1016/j.fertnstert.2004.11.064) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Immunoblotting of two-dimensional gel of PF. The proteins are separated in the first dimension by isoelectric focusing (pI, 4–7) and in the second dimension by molecular weight (10–180 kD). Polyvinylidene difluoride membranes were stained by using the AuroDye forte procedure; a rabbit anti-human Hp polyclonal antibody (Dako) was used as the primary antibody; the binding was detected using a horseradish peroxidase conjugated swine anti-rabbit polyclonal antibody and 3-amino-9-ethylcarbazole as substrate. The red line indicates the Hp β chains. Fertility and Sterility 2005 83, 1536-1543DOI: (10.1016/j.fertnstert.2004.11.064) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 (A) Hp β chain isoforms in a PF sample; 12 isoforms were observed in more than 75% of the PF and PL samples of the study population. (B) Expression of the 12 Hp β chain isoforms constantly present (>75% of cases) in the PF samples. (C) Expression of the 12 Hp β chain isoforms constantly present (>75% of cases) in the PL samples. Data are presented as mean ± SEM. Fertility and Sterility 2005 83, 1536-1543DOI: (10.1016/j.fertnstert.2004.11.064) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Expression (%Vo = normalized percent volume) of HpβE in the PF (A) and PL (B) of women with endometriosis and control subjects during the proliferative (P) and the secretory (S) phase of the menstrual cycle. Data are presented as mean ± SEM. Fertility and Sterility 2005 83, 1536-1543DOI: (10.1016/j.fertnstert.2004.11.064) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions