Key haplotypes in <i>MSRA</i> region showing association with MI in TSS cohort.

<p><b>^a</b>Numbers in parentheses indicate physical position on chr8 in base pairs (NCBI Build 36).</p><p><b>^b</b>All observed haplotypes with frequency >1% are shown; bold indicates haplotypes noted in text.</p><p>*Protective haplotypes that reached statistical significance after Bonferroni correction for all 2,890 haplotypes tested (<i>P</i><1.73×10⁻⁵).</p>‡<p>Near significant risk haplotype containing rs614197 (italicized), the SNP that was most highly associated with MI in the initial analysis.</p

Kaplan-Meier survival curves in CF mice according to <i>Msra</i> genotype.

<p>A. CF mice homozygous for a null <i>Cftr</i> allele (<i>Cftr</i>^−/−) and wild-type for <i>Msra</i> show high mortality due to intestinal obstruction around the time of weaning (ca. 21 days; n = 30). In contrast, survival is markedly improved in <i>Cftr</i>^−/− mice lacking one (n = 33) or two <i>Msra</i> alleles (n = 46) compared to wild-type (<i>P</i> = 0.022 and <i>P</i> = 0.0001, respectively; log-rank test). B. CF mice homozygous for a missense <i>Cftr</i> allele (<i>Cftr</i>^R117H/R117H) and wild-type for <i>Msra</i> display a low rate of mortality due to intestinal obstruction around the time of weaning (n = 14). Survival is not affected in <i>Cftr</i>^R117H/R117H mice lacking one (n = 51) or two (n = 51) <i>Msra</i> alleles compared to wild-type.</p

Population characteristics of GMS and NBS subjects.

<p>Population characteristics of GMS and NBS subjects.</p

Regional association of SNPs within a region of linkage to MI on chromosome 8p23.1.

<p>The inset plot shows the locus linked to MI on chromosome 8 identified by Blackman, et al <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002580#pgen.1002580-Blackman1" target="_blank">[12]</a>. The green shaded region under the peak extending from 17.9 cM (8.2 Mb) to 29.0 cM (17.2 Mb), where LOD score >1, indicates the region analyzed in the main plot. The map position of <i>MSRA</i> is denoted by an arrow at ∼20.2 cM. In the main plot, <i>P</i> values are plotted in log scale versus physical location in Mb. The SNP showing the strongest association with MI, rs614197, is represented by a diamond (<i>P</i> = 8.35×10⁻⁶). SNPs surrounding rs614197 are color coded to reflect their LD with this SNP (pair-wise r² using 1000 Genomes CEU, August 2009). The dashed line indicates the threshold for region-wide significance after Bonferroni correction for 2,896 SNPs (P<1.73×10⁻⁵). Genes, exon positions, and direction of transcription are denoted below plot (human genome build 18). Nine genes outside this interval were omitted for display purposes: <i>C8orf12</i>, <i>FAM167A</i>, <i>DEFB136</i>, <i>DEFB135</i>, <i>DEFB134</i>, <i>FAM66D</i>, <i>LOC392196</i>, <i>USP17L2</i>, and <i>FAM86B1</i>. The blue shaded region represents the 2 Mb encompassing rs614197 in which haplotype association was tested (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002580#pgen.1002580.s001" target="_blank">Figure S1</a>).</p

Patient characteristics.

<p><b>^a</b>Primary analysis included subjects from 133 “MI families” in which at least one sibling had MI.</p><p><b>^b</b><i>CFTR</i> mutation-specific analysis (i.e. p.Gly551Asp vs. p.Phe508del) utilized the entire TSS sample.</p

Haplotype association in <i>MSRA</i> region.

<p>In this schematic, haplotypes comprised of three consecutive SNPs are represented as dots connected by a line. Two overlapping haplotypes localized within intron 3 of <i>MSRA</i> had a statistically significant protective effect on MI (<i>P</i><1.73×10⁻⁵ after Bonferroni correction for 2,890 haplotypes tested in a 2 Mb region): rs10903323 T – rs4840475 G – rs17151637 A and rs4840475 G – rs17151637 A – rs6601427 C. A “risk” haplotype upstream of <i>MSRA</i>, rs586123 G – rs614197 G – rs2055729 C, was just below the threshold for significant association. Linkage disequilibrium patterns (pairwise r²) are displayed below the rs numbers corresponding to each SNP (physical location provided in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002580#pgen-1002580-t002" target="_blank">Table 2</a>).</p

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Basal feet polarity of brain ependymal cells and trachea epithelial cells was lost in the <i>Spag6</i>-deficient mice.

<p>Basal body images were taken with a transmission electron microscope. Notice that the basal feet point to the same orientation in the wild type animals (A: brain; C: trachea). However, they point to different orientation in the <i>Spag6</i>-deficient mice (B: brain; D: trachea). The number of basal body in the <i>Spag6</i>-deficient mice was significantly reduced in both brain and trachea. The arrows point to the basal feet. E. Average basal feet number counted from ten images randomly selected from each group. Horizontal lines represent means and SEMs. Three or four images were counted from each mouse, and three wild-type and three mutant mice were examined. * p<0.05. F. Circular plots of tracheal epithelial cell basal feet orientation in <i>Spag6</i>-deficient (left) and wild-type mice (right). For each mouse, basal foot orientation from five images was analyzed. For each image, the angel for one basal foot orientation was set as 0° (or 360°), angels of the rest basal feet were measured. Each plot represents the combined data from three mice as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107271#pone.0107271.s002" target="_blank">Figure S2</a> (* p<0.001 between the two groups).</p

Examination of rotational polarity of ciliary axoneme of brain ependymal cells and trachea epithelial cells by transmission electronic miscroscopy.

<p>Axoneme cross-sectional images were taken with a transmission electron microscope. The rotational polarity of each axoneme was evaluated by the angle of the line connecting the central pair. Notice that the orientation of the lines in wild type mice is similar (A: brain; C: trachea). while the orientation of the lines in the <i>Spag6</i>-deficient mice varies among axonemes (B: brain; D: trachea). E. Average axoneme number counted from ten images randomly selected from each group. Three or four images were counted from each mouse, and three wild-type and three mutant mice were examined. Horizontal lines represent the means and SEMs. * p<0.05.</p