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HeLa cells were cultured in DMEM (Sigma Aldrich) l supplemented with 5% fetal bovine serum, 2 mM - glutamine, 1% penicillin/streptomycin. Cells were trans- ferred to 24-well plates with 500 Al of regular growth medium/well the day before transfection. Transfections were performed with the Fugene 6 reagent as recommended by the manufacturer (Roche Diagnostics) with a mixture containing 1 Ag of each reporter plasmid and 2 ng of Renilla luciferase (Promega). Luciferase activity was measured after 24 h by using the Dual Luciferase Kit (Promega) in a Lumat LB9507 luminometer (EG&G Berthold). The luciferase activity of the reporter plasmids was normalized to the activity of Renilla luciferase. All transfections were performed in triplicate and repeated at least three times.

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Statistical analyses

VNTR allele frequencies in samples 1 and 2 were estimated by counting genes from the observed genotypes, after verification of Hardy–Weinberg equilibrium by allele shuffling (5000 random permutations). The null hypothesis of homogeneity between the genotypic (or allelic) pools of samples 1 and 2 was checked by permutation tests (1000 permutations). The statistical significance of the differences among the enhancer activity of different VNTR alleles in transfection experiments was tested by ANOVA and LSD post hoc test. The MATLAB statistical textbook was used for all the statistical analyses, excepted for linkage disequilibrium analysis between the markers G477T [15] and VNTR, for which the Arlequin software was used; this software utilizes a maximum likelihood estimation procedure by using geno- typic data from both homozygous and heterozygous subjects.