Summary of BDR by genotype of the SNP (rs295137) near <i>SPATS2L</i> with lowest P-value among all subjects in the primary populations.

<p>The TT genotype was associated with higher BDR (median 16.0; inter-quartile range (IQR) = [6.2, 32.4]), than the TC genotype (median 11.2; IQR = [5.2, 22.6]) or the CC genotype (median 10.3; IQR = [4.1, 21.7]).</p

Effect of siRNA-mediated <i>SPATS2L</i> knockdown on β₂-adrenergic receptor levels in HASM cells.

<p>(A) Knockdown efficiency of two <i>SPATS2L</i> siRNAs. Quantitative real-time PCR was done on RNAs extracted from HASM cells transfected with control non-targeting (NT) or <i>SPATS2L</i>-specific siRNAs. (B) Western blot analysis of β₂AR protein in <i>SPATS2L</i> knockdown HASM cells. Three independent experiments (siRNA transfection and Western blot analysis) were done in HASM cells. The upper panel is a representative of the three Western blots. Quantification of the β₂AR protein amount (normalized against the control β-actin protein) from three Western blots is shown in the lower panel.</p

Characteristics of GWAS subjects.

<p>Pre-BD = Pre-bronchodilator. SD = Standard deviation. For continuous variables, Mean (SD) and [Range] are shown.</p>*<p>Subjects were allowed to use rescue medication during wash-out period unless otherwise indicated.</p>**<p>550Kv3, 610, 317K, and 370K refer to Illumina BeadChips.</p>§<p>CARE trials included: CLIC, PACT, MARS.</p>¶<p>ACRN trials included: BAGS, DICE, IMPACT, PRICE.</p

Study overview.

<p>(A) Primary GWAS was conducted using subjects from CAMP, LOCCS, LODO, Sepracor, ACRN, and CARE cohort. Samples genotyped on Illumina platforms (i.e. CAMP/LOCCS/LODO/Sepracor) were pooled and analyzed first. Results from samples genotyped on Affymetrix platforms were analyzed separately and then combined to obtain the primary GWAS results. 1000GP imputed data was utilized to expand the primary GWAS association results. (B) Replication of the top (i.e. P-value<1E-04) SNPs from the primary GWAS were attempted in two independent populations: SARP and DAG. (C) The <i>SPATS2L</i> gene was selected for functional validation based on nominal replication of association results in SARP and analysis of publicly available resources.</p

Region of association near <i>SPATS2L</i> SNPs to BDR.

<p>The x-axis denotes position along Chromosome 2. The y-axis denotes −Log₁₀(P) corresponding to 1000GP imputed data P-values. LD between the SNP with the lowest P-value (rs295137) to each SNP in the plot is denoted in colors and was computed according to 1000GP June 2010 CEU data. Plot was created using LocusZoom <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002824#pgen.1002824-Pruim1" target="_blank">[44]</a>.</p

Primary GWAS Top Results (SNPs with Combined P-values <1e-05).

<p>CLLS = CAMP/LOCCS/LODO/Sepracor. CLLS used genotyped SNP data. CARE and ACRN used HapMap Phase 2 imputed data with Mach ratio of empirically observed dosage variance to the expected dosage variance (Rsq) values indicated.</p

Replication study of top SNPs in two independent populations (SARP and DAG).

*<p>rs295137 and rs295114 include SARP and DAG P-values. Remaining SNPs include SARP only.</p

Study overview.

<p>Mouse association study (in pink): (A) Baseline AHR slope measures were obtained for 31 mouse strains, and these were (B) used to perform an association study using EMMA software. (C) Mouse genes near or containing SNPs with p-values <0.001 were selected for follow-up study in human GWAS datasets. Integration of mouse results with human data (in blue): (D) HomoloGene maps were used to obtain human homologous genes corresponding to the top mouse genes. (E) Genes with SNPs having p-values <0.001 in the combined EVE meta-analysis or within race/ethnic specific GWAS were selected for (F) replication in other human GWAS.</p

Effect Sizes of Top <i>KCNIP4</i> SNP Associations.

<p>Odds ratios are shown for asthma (EVE combined cohort, GABRIEL, Sepracor/LOCCS/LODO/Illumina (SLLI), and DAG) and for AHR (SHARP AHR for change in LnPC20, DAG AHR for change in Ln(Slope)). EVE, GABRIEL, and SHARP include 95% confidence intervals. SLLI includes standard errors.</p

Top regions of association in two independent human asthma GWAS.

<p><i>KCNIP4</i> association in A) Sepracor/LOCCS/LODO/Illumina and C) GABRIEL. <i>PDZD2/GOLPH3/MTMR12/ZFR</i> association in B) Sepracor/LOCCS/LODO/Illumina and D) GABRIEL. The x-axis denotes position along corresponding human chromosome, while the y-axis denotes –Log₁₀(P). LD between the SNPs with the lowest P-value to other SNPs in the plots are denoted in colors and were computed according to HapMap Phase 2 CEU data using LocusZoom <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056179#pone.0056179-Pruim1" target="_blank">[55]</a>.</p