<i>PRRT2</i> coding variants identified in this study.

<p>Schematic diagram of the <i>PRRT2</i> gene and of the three PRRT2 protein isoforms. Mutation identified in this study are indicated in green (controls and HGDP), orange (controls, HGDP, and patients) and red (patients only).</p

<i>PRRT2</i> coding variants identified in individuals with ASD and controls.

a<p>Variants observed in the human genome diversity panel.</p>b<p>The p.A361_P362del was considered as probably damaging since it affects conserved amino acids of PRRT2.</p>c<p>Odds ratio, confidence intervals (CI) and P values were calculated only for populations from European ancestry using a 2-tailed Fisher exact test.</p

Chromatograms of the <i>PRRT2</i> A217PfsX8 mutation before and after whole genome amplification.

<p>The chromatograms of <i>PRRT2</i> sequence before (Native DNA) and after whole genome amplification of the DNA from three independent patients using two different protocols GenomiPhi DNA Amplification Kit (WGA-1) or Repli-G Whole Genome Amplification kit (WGA-2). The mutation was not present in the native DNA, but was detected after whole genome amplification.</p

Synonymous and nonsynonymous variations of <i>PRRT2</i> in worldwide populations.

a<p>Number of nonsynonymous and synonymous sites for PRRT2 are 668.5 and 231.5, respectively.</p>b<p>For the calculation of the pN/pS for Europe, the value of pS was set to 0.004 (the minimum of 1 synonymous variant observed in 159 individuals).</p

<i>PRRT2</i> variants identified in individuals from worldwide populations.

<p>A total of 961 individuals from the human genome diversity panel (HGDP) were sequenced for all <i>PRRT2</i> exons. The diameter of each circle is proportional to the number of individuals who were sequenced for <i>PRRT2</i>.</p

Genetic variability in Europe and Africa for all genes located within the 16p11.2 deletion.

<p>A. Nonsynonymous mutations per nonsynonymous sites (pN) and synonymous mutations per synonymous sites (pS) were estimated using the data from 4300 individuals from European ancestry and 2012 individuals from African ancestry available at the Exome Variant Server (<a href="http://evs.gs.washington.edu/EVS/" target="_blank">http://evs.gs.washington.edu/EVS/</a>). The horizontal lanes correspond to the means of pN and pS for the 27 genes. Difference of pN/pS between Europe and Africa were calculated using a 2-tailed Fisher exact test and the −Log₁₀ P value is indicated. B. Plot of the −Log₁₀ P values obtained for the difference between the pN/pS ratio in Africa and in Europe in relation the ratio (Europe pN/pS)/(Africa pN/pS).</p

Schematic illustration of the cellular interference mechanism associated with <i>PCDH19</i> mutations.

<p>A) In normal individuals, characterized by a homogeneous population of <i>PCDH19</i>-positive cells, neurons are able to form normal neuronal networks; B) In mutated male patients, hemizygosity leads to a homogeneous population of <i>PCDH19</i>-negative cells; in this condition, neurons preserve the ability to form normal neuronal networks; C) In heterozygous mutated females, random X inactivation leads to the co-existence of two <i>PCDH19</i>-positive and <i>PCDH19</i>-negative cell populations. These two cell populations cause divergent cell sorting and migration (due to attractive or repulsive interactions) and lead to abnormal neuronal networks. Somatic mosaicism in mutated males gives rise to the same pathological situation. The precise mechanisms by which the neuronal networks are altered are still unknown.</p

Detection of 9 different point mutations of <i>PCDH19</i> in 11 female patients by direct sequencing.

<p>A) Sequence electropherograms of the mutations and the missense variant (c.3319C>G/p.Arg1107Gly) identified in association with the c.859G>T/p.Glu287X nonsense mutation. The mutation nomenclature is based on the <i>PCDH19</i> transcript reference EF676096. Nucleotides are numbered according to the cDNA with +1 corresponding to the A of the ATG translation initiation codon in the reference sequence, according to the journal guidelines (<a href="http://www.hgvs.org/mutnomen" target="_blank">www.hgvs.org/mutnomen</a>). B) Alignment of the regions surrounding the mutations (indicated by an arrow) in orthologous and paralogous proteins, showing the high conservation of each affected amino-acid in vertebrates and in the delta protocadherin paralogous genes.</p

Clinical characteristics of patients with <i>PCDH19</i> mutations.

<p>2*: patient N 06 1258 is the sister of patient N 06 1257 (index case); PMD = psychomotor development, Nl = normal, F = febrile, unit = unilateral, GTC = generalized tonic-clonic, W-S = words-sentences, abs = absent, AED = anti epileptic drugs.</p

Pedigrees of the families and segregation analysis of the <i>PCDH19</i> deletion and point mutations.

<p>del/+, m/+ or v/+ denote individuals heterozygous for the deletion, mutation or variant, respectively; +/+ denotes individuals carrying homozygous wild-type alleles. Squares represent males, circles females; filled black symbols: patients diagnosed as having Dravet syndrome; right black half: Cognitive delay or impairment; left grey half: adolescence-onset idiopathic epilepsy. Dots in the middle of the squares indicate unaffected mutation carriers. The arrows indicate the index cases.</p