Additional file 2: Figure S2. of Toll-like receptors genes polymorphisms and the occurrence of HCMV infection among pregnant women

a and b Representative chromatograms of DNA fragments of various sequenced PCR products, encompassing TLR2 2258 G > A (A, B), TLR4 896 A > G (C, D), TLR4 1196 C > T (E, F) and TLR9 2848 G > A (G-I) SNPs. For TLR2 SNP, DNA forward strands were sequenced, and for TLR4 and TLR9—reverse strands were assayed. The numbers above some peaks of chromatograms indicate the following nucleotides determined in sequenced DNA fragments. Loci of the polymorphisms and genotypes analyzed in the study, are indicated with arrows. GG, GA, AA, AG, CT, CC—genotypes determined in studied TLR SNPs. (ZIP 1732 kb

DNA sequences, comprising polymorphic sites at <i>TLR4</i> (A, B) and <i>TLR9</i> (C-E) SNPs.

<p>Reverse strand sequences were determined for all the analyzed amplicons of <i>TLR</i> gene fragments; A. <i>TLR4</i> 896 A>G SNP; B. <i>TLR4</i> 1196 C>T SNP; C-E. <i>TLR9</i> 2848 G>A SNP. AA, CC, GG or GA—genotypes at described <i>TLR</i> SNPs.</p

PCR-RFLP analysis of genotypes at <i>TLR4</i> (A, B) and <i>TLR9</i> (C) SNPs.

<p>RFLP products were resolved in 2% agarose gels, stained with ethidium bromide; A. <i>TLR4</i> 896 A>G SNP; B. <i>TLR4</i> 1196 C>T SNP; C. <i>TLR9</i> 2848 G>A SNP. The numbers on the side of electropherograms indicate the lengths of separated DNA fragments. M—50 bp DNA marker; Ud—undigested PCR product; AA, AG, GG, GA, CC, CT or TT—genotypes at analyzed <i>TLR</i> SNPs.</p

Length of restriction fragments and genotypic profiles.

<p>^a SNP, single nucleotide polymorphism.</p><p>^b bps, base pairs.</p><p>Length of restriction fragments and genotypic profiles.</p

Primer sequences, annealing temperatures and amplicon lengths, obtained in multiplex nested PCR assay for SNPs in the <i>TLR4</i> and <i>TLR9</i> genes.

<p>^a No., number.</p><p>^b SNP, single nucleotide polymorphism.</p><p>^c bps, base pairs.</p><p>Primer sequences, annealing temperatures and amplicon lengths, obtained in multiplex nested PCR assay for SNPs in the <i>TLR4</i> and <i>TLR9</i> genes.</p

Visualization of PCR-RFLP products for <i>TLR3</i> SNP (A) and <i>TLR7</i> SNP (B) genotyping.

<p>Gel image: A) rs3775290: 1. wild type CC, 2. heterozygous CT, 3. homozygous recessive TT; rs3775291: 4. wild type CC, 5. heterozygous CT, 6. homozygous recessive TT; rs3775296: 7. wild type CC, 8. heterozygous CA, 9. homozygous recessive AA. B) rs179008: 1. wild type AA, 2. heterozygous AT, 3. homozygous recessive TT; rs5741880: 4. wild type GG, 5. heterozygous GT, 6. homozygous recessive TT. Alignment markers (15 bp, 1 kbp).</p

The distribution of allele frequencies of TLR SNPs in infants with and without HCMV infection.

<p>The distribution of allele frequencies of TLR SNPs in infants with and without HCMV infection.</p

Selected single-nucleotide polymorphisms in the <i>TLR3</i> and <i>TLR7</i> genes.

<p>Selected single-nucleotide polymorphisms in the <i>TLR3</i> and <i>TLR7</i> genes.</p

Frequency and distribution of <i>TLR</i> genotypes and their association with HCMV infection.

<p>Frequency and distribution of <i>TLR</i> genotypes and their association with HCMV infection.</p

Restriction enzymes and length of the restriction fragments.

<p>Restriction enzymes and length of the restriction fragments.</p