Sequencing of <i>Dclre1c-</i>gene.

<p>(A) Two heterogeneous and four homogeneous mutant mice were analyzed using next-generation-sequencing techniques. Displayed is the coverage of the bases in exons 9 to 12. (B) Graphical representation of the genomic organization of <i>Dclr1c</i> between exons 9 and 12. Exons are indicated by black boxes, while arrows indicate primer binding-sites. The upper row represents the organization in the wild-type genome while the lower row represents the gene in the mutant. (C) Results of PCR reactions with the indicated primer sets. DNA was visualized on ethidium bromide agarose gels. (D) Primer p3f and p8r were used to amplify DNA from mutant animals and the purified PCR products were used for Sanger DNA-sequencing. The resulting sequences were aligned to the <i>Dclre1c</i>-sequence according to the “Gene” database of NCBI. (*) indicates matching bases, (-) indicates gaps in the alignment. Shown is the 5´ beginning of the deletion (before //) as well as the 3´ end of the deletion (after //).</p

Single nucleotide polymorphism (SNP) analysis of mutant mice.

<p>(A) Mutant mice on C57BL/6 background were crossed to C3H and the offspring (F1) was used for an intercross yielding F2 animals. Blood of these animals was analyzed for the percentage of T cells (TCR-β ⁺ DX5^-) and B cells (CD19⁺MHCII⁺). F2 animals were grouped according to the presence (unaffected) or absence (affected) of T and B cells and affected animals were used for SNP-analyses. (B) Graphical representation of SNP distribution in randomly selected F2 and control animals. SNPs derived from C57BL/6 are in blue, SNPs from C3H are in red and heterogenous SNPs are yellow. Shown are only informative SNPs at the beginning of chromosome 2. The chip contained 377 SNP of which 244 were informative in our cross. In total we performed SNP-analyses from 57 F2 animals as well as from 6 F1, C57BL/6 and C3H control animals each.</p

Initial characterization of cell subsets in the spleens of mutant mice.

<p>(A) Single cell suspensions of spleens were stained for B cells (CD19⁺B220⁺) or T cells (CD4⁺ or CD8⁺), which were further analyzed for expression of CD3 (lower panel). Cells were gated on live lymphocytes. (B) Single cell suspensions of spleens were stained with the indicated antibodies to identify DC (CD11c⁺MHCII⁺), NK cells (NK1.1 ⁺ CD3^-), granulocytes (Gr1 ⁺ CD3^-) and CD11b⁺ monocytes. FACS-plots and statistics are representative results from one out of six experiments with similar outcome (n=3 mice per group). Bar graphs represent mean number of live cells ± SD in the respective gate.</p

ANOVA results (comparison children with ADHD versus healthy control children).

<p>ANOVA results (comparison children with ADHD versus healthy control children).</p

Group effects and effects of stimulant medication.

<p>Grand average of lateralized motor potentials ([C3<b>−</b>C4]/2) for children with ADHD and healthy control children. In ADHD children, the potential waveform before and after intake of 10 mg methylphenidate are shown – the order in which the sessions were recorded was counterbalanced.</p

Scatterplot showing the correlation between response speed and lateralized initial motor potential peak amplitude (iMP’) for children with ADHD and healthy control children ([C3−C4]/2).

<p>Scatterplot showing the correlation between response speed and lateralized initial motor potential peak amplitude (iMP’) for children with ADHD and healthy control children ([C3−C4]/2).</p

Time-course and topography of response-locked movement-related potentials (initial movement related potential peak - iMP) including the topography of iMP lateralization ([C3−C4]/2).

<p>For control children, averages of all responses are illustrated together with a separate presentation of averages of fast (below median reaction time) and slow responses (above median reaction time). For children with ADHD, responses on and off methylphenidate are presented. For effects of response speed in ADHD see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039012#pone-0039012-g004" target="_blank">Figure 4</a>. Note how the rather symmetrically distributed stimulus-related P300 shadowed MRP in the topography before the subtraction of symmetrically distributed potential components by the calculation of lateralization. iMP time-course (thick black arrows) and lateralized topography around C3 (grey arrows) are in good agreement with previous literature. mPINV (lateralized negativity at C3 in the time interval 500–800 ms) was not shadowed by P300 any more. Its topography is presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039012#pone-0039012-g002" target="_blank">Figure 2</a>.</p

Lateralized motor potential (iMP’) and motor postimperative negative variation (mPINV’) amplitudes for the ADHD and the healthy control group ([C3<b>−</b>C4]/2).

<p>Lateralized motor potential (iMP’) and motor postimperative negative variation (mPINV’) amplitudes for the ADHD and the healthy control group ([C3<b>−</b>C4]/2).</p

Motor PINV topography for healthy control children.

<p>(top; from left to right: all responses, fast responses below median reaction time, slow responses above median reaction time) <b>and children with ADHD</b> (bottom; from left to right: responses off and on methylphenidate) for the motor PINV time interval (500–800 ms after the response trigger, motor post-processing).</p

ANOVA results (effects of methylphenidate).

<p>ANOVA results (effects of methylphenidate).</p