Activity of IL-33 variants in human CD4 T cell bioassay.

<p>Purified human blood CD4 T cells were incubated in the presence or absence of IL-2 and IL-12 for 72 hours and IFN-γ was measured in the cell-free supernatants (as described in online methods). Both IL-33 (95–270) (Amgen) and IL-33(112–270) (R&D Systems) were used as positive controls and two independent lots of IL-33(95–204) were tested. (<b>A</b>) IL-33, IL-2 and IL-12 dependence of assay with IL-33 at 10ng/mL; (<b>B</b>) Dose titration of IL-33 variants (in the presence of 10 ng/mL IL-2 and 10 ng/mL IL-12). (<b>C</b>) ST2 dependence of response. IL-33 variants (10 ng/mL plus IL-2 and IL-12) were tested in the presence of 25 ug/mL human IgG2 control antibody or IgG2 anti-human ST2 blocking antibody.</p

Association of common sequence variants in <i>IL33</i> and <i>IL1RL1</i> with eosinophil counts and asthma in Iceland.

<p>Association of common sequence variants in <i>IL33</i> and <i>IL1RL1</i> with eosinophil counts and asthma in Iceland.</p

RNA analysis of <i>IL33</i> mRNA expression and carrier status of the splice acceptor variant rs146597587.

<p><b>(A)</b> Gene structure of <i>IL33</i> and location of rs146597587 SNP that changes a canonical splice acceptor site from AG to AC. The mutation lies at the junction of coding exons 6 and 7. Exon 7 begins LHKCE and encodes a significant portion of the cytokine domain. Also, indicated is the location of an elastase cleavage site at amino acid 95 that releases the cytokine domain from the N terminus prodomain and leads to a more active cytokine. <b>(B)</b> Expression in FPKM of <i>IL33</i> in adipose tissue for rs146597587 non-carriers (GG, N = 744) and carriers (GC, N = 6) based on RNA-Seq data; <i>P(Wilcox)</i> = 0.0015, median non-carriers: 34.1, carriers: 21.1 (i.e. heterozygotes have 38% lower expression than non-carriers). <b>(C)</b> Ratio of RNA-Seq read coverage in last intron of <i>IL33</i> versus its last exon (splice acceptor resides at this intron-exon boundary) for rs146597587 non-carriers (GG, N = 738) and carriers (GC, N = 9); <i>P</i>(Wilcox) = 2.5×10^–7, mean non-carriers: 0.67%, carriers: 9.0%. <b>(D)</b> Proportion of RNA-Seq read with ALT allele for the synonymous variant rs10975519 (MAF = 29%) for rs146597587 non-carriers (GG, N = 309) and carriers (GC, N = 5); a chromosome carrying rs146597587-C carries the ALT allele of rs10975519; <i>P</i>(Wilcox) = 1.4×10^–4, mean non-carriers: 0.48, non-carriers: 0.20.</p

Conditional analysis for eosinophil counts associations in the region around <i>IL1RL1</i>.

<p>Plot shows an 800kb overview around the <i>IL1RL1</i> gene on chromosome 2. Black circles (o) show-log₁₀ <i>P</i> as a function of hg38 coordinates for unadjusted associations with eosinophil counts; red crosses (+) correspond to eosinophil counts associations after adjusting for the variant rs13020553; blue ‘x’ symbols correspond to eosinophil counts associations after adjusting for both rs13020553 and rs6719123. The position of the two variants rs13020553 and rs6719123 are indicated by vertical broken lines. Genes are shown in blue and recombination rates are reported in cM/Mb.</p

Blood eosinophil counts from wild type and <i>il33</i>-deficient mice.

<p>Blood were collected from male and female 10–12 week old mice and eosinophils were enumerated as described in the online methods. The horizontal bars indicate means within groups. Significance calculated using the unpaired T test, p = 0.014 males, p = 0.0001 females.</p

Overview of eosinophil counts associations in the region around <i>IL33</i>.

<p><b>(A)</b> shows an 800kb overview centered on <i>IL33</i> on chromosome 9 and (<b>B)</b> shows a 100kb overview around the <i>IL33</i> gene. Black circles show -log₁₀ <i>P</i> as a function of hg38 coordinates for associations with eosinophil counts and red crosses correspond to eosinophil counts associations after adjusting for the three variants rs2095044, rs146597587 (splice acceptor) and rs10758750 that are indicated by vertical broken lines in (<b>B)</b>. Genes are shown in blue and recombination rates are reported in cM/Mb. (See <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006659#pgen.1006659.s002" target="_blank">S1 Fig</a> for intermediate results from stepwise regression.)</p

Additional file 3: of Exome-chip meta-analysis identifies novel loci associated with cardiac conduction, including ADAMTS6

Video S1. (Quicktime) Video to illustrate the DORV phenotype finding in an Adamts6 mutant heart. (MOV 1983 kb

Additional file 2: of Exome-chip meta-analysis identifies novel loci associated with cardiac conduction, including ADAMTS6

Figure S1. Manhattan plot for European and African-American ancestry single variant analysis. Figure S2. Quantile-quantile plot for European and African-American ancestry single variant analysis. Figure S3. Manhattan plot for EA single variant analysis. Figure S4. QQ plot for EA single variant analysis. Figure S5. Manhattan plot for AA single variant analysis. Figure S6. Quantile-quantile plot for AA single variant analysis. Figure S7. Miami plot European and African-American ancestry sex-stratified single variant analysis. Figure S8. Quantile-quantile plots for European and African-American ancestry sex-stratified single variant analyses. Figure S9. Normal morphology of adult Adamts6 heterozygous hearts. (DOCX 4290 kb