Comparative analysis of the seminal plasma proteomes of oligoasthenozoospermic and normozoospermic men  Elisa Giacomini, Blendi Ura, Elena Giolo, Stefania Luppi, Monica Martinelli, Rodolfo C. Garcia, Giuseppe Ricci  Reproductive BioMedicine Online  Volume 30, Issue 5, Pages 522-531 (May 2015) DOI: 10.1016/j.rbmo.2015.01.010 Copyright © 2015 Reproductive Healthcare Ltd. Terms and Conditions

Figure 1 Two-dimensional separation of seminal plasma proteins. Typical seminal plasma protein separation by isoelectric focusing (pI 3 to 11) followed by sodium dodecyl sulphate polyacrylamide electrophoresis. Major protein constituents are shown encircled. pI, isoelectric point. Reproductive BioMedicine Online 2015 30, 522-531DOI: (10.1016/j.rbmo.2015.01.010) Copyright © 2015 Reproductive Healthcare Ltd. Terms and Conditions

Figure 2 (2A) Protein spots for which there was a statistically significant difference in intensity between idiopathic oligoasthenozoospermia and normozoospemia. Spots of intensity differing by more than 1.5-fold (average, P < 0.05, analysis of variance) on comparison of 10 men with idiopathic oligoasthenozoospermia with 10 men with normozoospermia are encircled in red. Intensities are greater in normozoospermia than in idiopathic oligoasthenozoospermia (number 200, 210, 504, 515) or greater in idiopathic oligoasthenozoospermia than in normozoospermia (number 481, 663), as shown in 2B; (2B) the comprehensive total of spots is 1028. Intensities (normalized volume, y axis of graphs) of each of the protein spots on all gels, 10 of normozoospermia and 10 of idiopathic oligoasthenozoospermia, are shown. The statistical significance values (analysis of variance) of the differences are indicated. ANOVA = analysis of variance; iOA = idiopathic oligoasthenozoospermia; N = normozoospermia; pI = isoelectric point. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) Reproductive BioMedicine Online 2015 30, 522-531DOI: (10.1016/j.rbmo.2015.01.010) Copyright © 2015 Reproductive Healthcare Ltd. Terms and Conditions

Figure 3 Western blot enhanced chemiluminescence detection and quantification of NPC2 (epididymal secretory protein E1) in seminal plasma from four men with normozoospermia and four men with idiopathic oligoasthenozoospermia. The image shown corresponds to a film exposure of 2 min. The intensity of the bands was quantified using ImageJ (NIH, USA). Mean intensities and standard deviations are indicated for each group (idiopathic oligoasthenozoospermia and normozoospermia) by vertical bars. The ratio of mean intensities in idiopathic oligoasthenozoospermia compared with normozoospemia was 2.67. The differences between idiopathic oligoasthenozoospermia and normozoospermia, indicated by an asterisk, are statistically significant (P = 0.034, t-test). iOA = idiopathic oligoasthenozoospermia; N = normozoospermia. Reproductive BioMedicine Online 2015 30, 522-531DOI: (10.1016/j.rbmo.2015.01.010) Copyright © 2015 Reproductive Healthcare Ltd. Terms and Conditions