Analysis of chicken plasma IgT binding CLL.

<p>Per fragment (size kDa) the average extinction (least square means) per line of hens is shown, with standard error (SE) and P-value.</p>1<p>Line: Hg = genotypic high hens (N = 12), Cg = control hens (N = 11), Lg = genotypic low hens (N = 11), Ch = phenotypic high hens (N = 12), Cl = phenotypic low hens (N = 12).</p><p>Extinctions within a row with different superscripts^a,b,c differ significantly (P<0.05).</p>*<p>P<0.05,</p>**<p>P<0.01,</p>***<p>P<0.001.</p

Western Blots representing the same set of birds for different isotypes.

<p>From top to bottom: A) IgTotal, B) IgM and C) IgG. Lanes 1–4: genotypically low (Lg) hens, 5–10: phenotypically high (Ch) hens, C = positive control (rooster plasma), m = marker, and other lanes are negative control (dilution fluid).</p

Correlation of staining patterns of all bands stained by all isotypes (A) or stained by IgTotal (B), IgM (C), and IgG (D) autoantibodies to CLL.

<p>Chickens were one year of age and divergently selected for 29 generations for High (red circles: Hg), Low (green diamonds: Lg), and Control (purple squares: Cg) agglutination titres to SRBC at 5 weeks of age, and C line birds phenotypically selected for high (yellow bars: Ch) or low (blue bars: Cl) NAb levels to KLH at 16 weeks of age. Ordination plots by PCA. Graphs include 11–12 birds from each line as also used for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072276#pone-0072276-g001" target="_blank">Figures 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072276#pone-0072276-g002" target="_blank">2</a>. Only CLL fragments that showed significant line differences for staining intensity (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072276#pone-0072276-t004" target="_blank">Tables 4</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0072276#pone-0072276-t006" target="_blank">6</a>) are shown. ‘Eigenwaardes’ were 12.8% horizontal and 11.1% vertical axes (A), 17.9% horizontal and 11.6% vertical axes (B),16.5% horizontal and 15.4% vertical axis (C), and 20.8% horizontal and 12.5% verical axis (D), respectively. Numbers in <i>Ithalic</i> represent CLL fragments most informative for line clustering.</p

Analysis of chicken plasma IgG binding CLL.

<p>Per fragment (size kDa) the average extinction (least square means) per line of hens is shown, with standard error (SE) and P-value.</p>1<p>Line: = genotypic high hens (N = 12), Cg = control hens (N = 11), Lg = genotypic low hens (N = 11), Ch = phenotypic high hens (N = 12), Cl = phenotypic low hens (N = 12).</p><p>Extinctions within a row with different superscripts^a,b,c differ significantly (P<0.05).</p>*<p>P<0.05,</p>**<p>P<0.01,</p>***<p>P<0.001.</p

Least square means plus standard error (SE) of immunoglobulin M (IgM) and immunoglobulin G (IgG) titers and titers of total antibodies (IgTotal) binding chicken liver lysate (CLL).

a,b,c<p>Lines differ significantly (<i>P</i><0.05) when different superscript.</p>1<p>Hg = genotypic high hens, Cg = control hens, Lg = genotypic low hens, (N = 15 birds/line).</p>2<p>microgram/ml coating of CLL to ELISA plates.</p>3<p>Line effect:</p>***<p>P<0.001.</p

Average number of antibody recognized CLL bands per chicken, without correcting for the different lines.

<p>Average number of antibody recognized CLL bands per chicken, without correcting for the different lines.</p

Graphic representation of Western Blots.

<p>On each figure the sample data from one phenotypically high (Ch) hen is shown for the three different isotypes. From top to bottom; IgTotal, IgM and IgG.</p

Analysis of chicken plasma IgM binding CLL.

<p>Per fragment (size kDa) the average extinction (least square means) per line of hens is shown, with standard error (SE) and P-value.</p>1<p>Line: Hg = genotypic high hens (N = 12), Cg = control hens (N = 11), Lg = genotypic low hens (N = 11), Ch = phenotypic high hens (N = 12), Cl = phenotypic low hens (N = 12).</p><p>Extinctions within a row with different superscripts^a,b,c differ significantly (P<0.05).</p>*<p>P<0.05,</p>**<p>P<0.01,</p>***<p>P<0.001.</p