Polymorphisms of nucleotide-excision repair genes may contribute to sperm DNA fragmentation and male infertility Aihua Gu, Guixiang Ji, Yong Zhou, Yan.

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Polymorphisms of nucleotide-excision repair genes may contribute to sperm DNA fragmentation and male infertility Aihua Gu, Guixiang Ji, Yong Zhou, Yan Long, Xiangguo Shi, Guangbo Fu, Shoulin Wang, Ling Song, Xinru Wang Reproductive BioMedicine Online Volume 21, Issue 5, Pages 602-609 (November 2010) DOI: 10.1016/j.rbmo.2010.06.025 Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions

Figure 1 Detection of sperm DNA damage using Tdt (terminal deoxynucleotidyl transferase)-mediated dUTP nick end labelling assay. Histogram data from flow cytometry shows (A) a negative control, (B) one sample with 24.9% spermatozoa (M2) labelled with strong fluorescein isothiocyanate fluorescence and (C) distribution of sperm DNA damage in 620 ejaculates. The x axis in A and B: the percentage of fluorescence intensity measured in FL1-H channel. Reproductive BioMedicine Online 2010 21, 602-609DOI: (10.1016/j.rbmo.2010.06.025) Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions

Figure 2 Association between XPA(–4) genotypes and the values of sperm DNA fragmentation, sperm concentration and sperm motility. The box-and-whisker plots depict the distribution of the transformed (A) TUNEL value, (B) sperm concentration and (C) motility among 620 subjects with the different genotypes. The boxes represent the 25th and 75th percentiles; whiskers cover the extent of the data on 1.5 interquartile range. The median value is denoted as the line that bisects the boxes. Circles represent the outlier values. Significant difference measured by multiple linear regression. ∗P<0.05 compared with the GG groups. Reproductive BioMedicine Online 2010 21, 602-609DOI: (10.1016/j.rbmo.2010.06.025) Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions

Figure 3 Effect of the XPA(–4) A/G polymorphism on the transcriptional activity. (A) Schematic representation of reporter plasmids containing the –4 G or –4 A allele, which was inserted upstream of the luciferase reporter gene in pGL3 basic plasmid. (B) DNA sequencing results of reporter plasmids containing the –4 G or –4 A allele; arrows indicate the positions of nucleotide changes. (C) The two constructs were transiently transfected into the MCF-7, HeLa and COS-7 cells, respectively. The luciferase activity of each construct was normalized against the internal control of renilla luciferase. The data indicate the mean values with the standard deviation from three independent experiments. ∗P<0.05 compared with the construct counterpart. Reproductive BioMedicine Online 2010 21, 602-609DOI: (10.1016/j.rbmo.2010.06.025) Copyright © 2010 Reproductive Healthcare Ltd. Terms and Conditions