The extent of inbreeding depression as a function of deleterious mutation rate.

<p>Mean inbreeding depression under complete outcrossing (<i>S</i> = 0; solid line) and complete selfing (<i>S</i> = 1; broken line), were obtained analytically for an infinite population (equations 1a, b). The dashed-dotted line shows the threshold level of inbreeding depression ( = 0.5), below which selfing is evolutionarily stable. Selection parameters are set to be <i>h</i> = 0.2 and <i>s</i> = 0.1, based on available data for <i>Drosophila melanogaster </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033541#pone.0033541-Mukai1" target="_blank">[19]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033541#pone.0033541-Crow1" target="_blank">[20]</a>.</p

The rate of mutation accumulation as a function of population size.

<p>For each combination of <i>U</i> ( = 0.1, 0.3, and 0.5) and <i>s</i> ( = 0.01 and 0.1), simulation results are illustrated for populations under complete selfing (<i>S</i> = 1; broken lines), predominant selfing with slight outcrossing (<i>S</i> = 0.97; heavy solid lines), or complete outcrossing (<i>S</i> = 0; thin solid lines). The average rate of mutation accumulation was obtained by taking the arithmetic mean over 1000 generations after the systems reached steady states. Error bars indicate the standard deviations. Throughout, the degree of dominance was <i>h</i> = 0.2. The data points for situations with no fixation (i.e. when the mutation accumulation rate was zero) were omitted from the figure. Note that ordinates are given in (common) logarithmic scale. Note also that the ranges of parameters shown in the two axes vary among panels.</p

Comparison of the mutation accumulation rates when deleterious mutation rates vary between mating systems.

<p>Simulation results are illustrated for populations under complete selfing (<i>S</i> = 1; broken lines), predominant selfing with slight outcrossing (<i>S</i> = 0.97; heavy solid lines), or complete outcrossing (<i>S</i> = 0; thin solid lines). The selection coefficient was either <i>s</i> = 0.01 (A, B) or 0.1 (C, D), with <i>h</i> = 0.2 in all cases. Distinct rates of deleterious mutation were assigned to selfing and outcrossing populations, for which appropriate mutation rates were predicted from the analytical model as developed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033541#pone.0033541.s001" target="_blank">Text S1</a>. The mutation rate was kept at <i>U</i> = 0.1 (A, C) or 0.3 (B, D) for populations with complete selfing or slight outcrossing, whereas <i>U</i> = 0.5 for completely outcrossing populations. Error bars indicate the standard deviations. Note that ordinates are given in (common) logarithmic scale. The data points for situations with no fixation (i.e. when the mutation accumulation rate was zero) were omitted from the figure. Note also that the range of population size shown in the horizontal axis varies among panels.</p

SIV antigen-specific CD8⁺ T-cell responses.

<p>SIV Gag-, Pol-, Vif-, Vpx/Vpr-, Tat-, Rev-, Nef-, and Env-specific CD8⁺ T-cell responses were measured by detection of antigen-specific IFN-γ induction using PBMCs at weeks 26-30 post-challenge.</p

Binding properties of IgGs to SIV antigens.

<p>The non-NAb cocktail, ten non-NAb IgG lots derived from ten macaques, and CAb were subjected to immunoblotting (ZeptoMetrix). A representative result from two experiments is shown.</p

Predominant nonsynonymous env mutations.

<p>Viral cDNAs encoding Env were amplified from plasma RNAs obtained at 7-9 months (R10-001, R10-004, and R06-029) or 12 months (other animals) and subjected to sequencing analysis. Amino acid substitutions are shown. The asterisk (*) represents a deletion and the double asterisk (**) represents coexistence of multiple deletion patterns. </p

ADCVI activity of the non-NAb cocktail and non-NAb IgG lots.

<p>The reduction in SIV p27 concentration in the supernatant from SIV-infected cell culture with non-NAbs compared to that without antibodies is shown. A representative result, means of duplicate samples, from two experiments is shown.</p

Binding properties of IgGs to SIV virions.

<p>Polyclonal IgGs purified from macaque plasma were subjected to whole virus ELISA using purified SIV_mac239 virions as the antigen. Results on ten IgG lots derived from ten macaques without detectable neutralizing activity (non-NAbs; black lines), five with neutralizing activity (NAbs; red), and a control IgG (CAb; green) are shown in the left panel. Results on the non-NAb cocktail and three non-NAb lots composing the cocktail are in the right. The dotted line represents background absorbance. Time points of plasma sampling are shown in parentheses following the macaque IDs. A representative result, means and SDs of duplicate samples, from two experiments is shown.</p

Expression of the CaMKIIβ throughout the brain by AAV-PHP.eB.

Volume-rendered and single-plane images of the brain expressing H2B-mCherry under hSyn1 promoter by the AAV (mCherry, green) counterstained with RD2 (red). A volume-rendered image is shown in the center. Single-plane and magnified images are shown for cerebral cortex, thalamus, hippocampus, midbrain, cerebellum, striatum, and olfactory bulb. Scale bar in the center image, 3 mm; other scale bars, 100 μm. AAV, adeno-associated virus; CaMKIIβ, calmodulin-dependent protein kinase IIβ; hSyn1, human synapsin-1. (TIFF)</p

Robust sleep induction by CaMKIIβ T287D mutant.

(A) Expression levels of endogenous CaMKIIβ and AAV-mediated transduced CaMKIIβ in the brain. Camk2bFLAG/FLAG represents homo knock-in mice in which the FLAG tag was inserted into the endogenous Camk2b locus. PBS: PBS-administrated mice. Immunoblotting against FLAG-tagged protein indicates that AAV-mediated expression of CaMKIIβ is lower than the expression level of endogenous CaMKIIβ. (B) Calculated transduction efficiency plotted against sleep duration. Transduction efficiency is an estimation of the number of AAV vector genomes present per cell in a mouse brain. After the SSS measurements, we purified the AAV vector genomes from the mice brains and then quantified them with a WPRE-specific primer set and normalized to mouse genomes. (C) Sleep transition profiles of mice expressing CaMKIIβ T287-related mutants shown in Fig 1F. The shaded areas represent SEM. (D) Sleep parameters during light or dark period of mice expressing CaMKIIβ T287-related mutants shown in Fig 1F. Multiple comparison tests were performed between all individual groups in each phase. (E, F) Sleep/wake parameters of mice expressing S114-related CaMKIIβ mutants (C) and S109-related CaMKIIβ mutants (D), averaged over 6 days. The shaded areas represent SEM. Multiple comparison tests were performed between all individual groups and resulted in no significant differences. The underlying numerical data can be found in S1 Data, and uncropped or raw image files for S3A Fig are provided in S2 and S3 Data files. Error bars: SEM, *p p p (PDF)</p