Associations between CD14 C-159T and TLR4 Asp299Gly variants with IMD using different genetic models.

<p>Odds ratios (OR) and 95% confidence intervals (CI) were obtained from univariate logistic regression analysis. Due to minor allele frequency of the 299Gly allele of TLR4, no recessive genetic model was applied.</p

<i>POMC</i> CpG methylation pattern of CpG island 1 and 2.

<p>Analysis example of the DNA methylation pattern of human peripheral blood cells (PBC) from a normal weight individual after sequencing of 10 different clones of the PCR amplification product. Filled cycles represent methylated CpGs, open cycles non-methylated CpG positions, which are numbered according to their relative position to the start of the next exon. Alu element positions and P300 binding site are marked.</p

<i>POMC</i> intron2 exon3 genomic sequence, P300 ChIP assay, real-time PCR analysis, and real-time <i>POMC</i> PCR.

<p>(A) <i>POMC</i> intron2 exon3 genomic sequence with annotated ChIP primer localization and P300 binding site. (B) P300 ChIP assay was performed at three independent occasions within human peripheral blood cells. <i>POMC</i> fragment was amplified with two different primer pairs (fragment 1 and 2). Confirmation of a published P300 binding site within the insulin gene promoter in β-TC3 cells was used as a positive control <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002543#pgen.1002543-Chakrabarti1" target="_blank">[34]</a> (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002543#pgen.1002543.s005" target="_blank">Figure S5</a>). (C) Real time PCR analysis (PCR fragment 2) of the P300 ChIP results of 4 obese patients with a hypermethylation variant and 6 individuals with hypomethylated intron 2-exon 3 intersection at <i>POMC</i> CpG island 2. (D) Real-time <i>POMC</i> PCR (<i>POMC</i> exon 2–3 and <i>POMC</i> exon3) of cDNA extracted from PBC of hypomethylated normal weight individuals (n = 20), hypermethylated obese patients (n = 20), and obese individuals without the hypermethylated variant (n = 20) reveal reduced POMC gene expression in the hypermethylated samples.</p

Meningococcal disease, age of infection and serogroup distribution.

<p>Numbers of cases are plotted against age of infection, presented as age groups in a 2-year interval, with respect to serogroups. Black bars indicate all cases, grey bars indicate serogroup B, and white bars indicate serogroup C.</p

Longitudinal analysis of <i>POMC</i> DNA methylation score (%) (CpG position −4 to +6) before and after the development of obesity in altogether 21 participants of the MAS birth cohort.

<p>Each line represents one individual course of DNA methylation score prior to and after weight gain.</p

<i>POMC</i> DNA methylation pattern of CpG island 1 and 2 in normal-weight adolescents (n = 12), laser-microdissected MSH positive cells of the human arcuate nucleus (n = 5) and human newborn samples (n = 8).

<p>CpG positions are numbered according to their relative position to the next exon start.</p

The DNA methylation was analysed with two different bisulphite based protocols in two independent cohorts.

<p>A, B, C agarose-embedded DNA was analysed by bisulphite sequencing. In D, E, F DNA was bisulphite treated in a non-agarose-bead protocol. A/D Diagram of the DNA methylation score (%) of <i>POMC</i> CpG island 2 within PBC of normal weight (red) and obese individuals (blue) (p<0,001). CpG positions are numbered according to their relative position to the next exon start. B/E Box plots analysis represents the statistic differences from the mean CpG methylation score (%) of CpG position −4 to +6 in normal weight individuals versus obese patients. C/F Percent of normal weight (red) and obese (blue) individuals, who showed DNA methylation at the annotated single CpG position (−1 to +5) in relation to the analysed number of samples.</p

Questionnaire items used for scoring.

<p>Numbers in brackets (%) are frequencies of parents' answers, denominator of n = 1226 in Leicester subsample used for generating the tool, and of n = 140 in MAS-90 subsample. * before/after imputation with values from prior follow-up assessments and baseline.</p><p>Questionnaire items used for scoring.</p

Comparison of study characteristics.

<p>Comparison of study characteristics.</p

Predicted probability of developing asthma 5 years later, for a single particular score value.

<p>Performance measures by score in table above. Primary inclusion, scoring, and outcome definitions. Probabilities from original paper (dotted line). * Values not meaningful and/or no accurate estimate possible. PPV/NPV, positive/negative predictive value; LR+/-, positive/negative likelihood ratio.</p