Additional file 1: of Loss of maternal EED results in postnatal overgrowth

Supplementary information. (PDF 389 kb

Additional file 5: Table S4. of Multiallelic copy number variation in the complement component 4A (C4A) gene is associated with late-stage age-related macular degeneration (AMD)

Sensitivity analysis for C4A copy number association. (DOCX 17 kb

Additional file 6: Figure S2. of Multiallelic copy number variation in the complement component 4A (C4A) gene is associated with late-stage age-related macular degeneration (AMD)

Subgroup/sensitivity analysis in the pooled study of C4A copy number. Odds ratios and corresponding 95 % confidence intervals are given with the size of each rectangle representing the respective relative number of cases and controls in each subgroup. The protective effect of increasing C4A copy number is stronger in females and increases with age. Both effects can also be observed when conditioning on known AMD associated risk variants at the C2/CFB locus (rs429608, rs114190211, rs204993 and rs142511358 [9]) and are present in each of the individual studies. 95 % confidence intervals are indicated; N stands for the total number of individuals included in the analysis. (TIF 1946 kb

Additional file 9: Figure S4. of Multiallelic copy number variation in the complement component 4A (C4A) gene is associated with late-stage age-related macular degeneration (AMD)

AMD associated haplotypes at the C2/CFB locus on chromosome 6. The phase at C2/CFB was assessed with SHAPEIT2 for each individual. In total, we found six haplotypes (H1-H6) with an allele frequency ≥ 1 % in the study. The haplotypes are characterized by the presence or absence of AMD associated alleles from four inpendent variations at this locus (rs429608, rs114190211, rs204993 and rs142511358 [9]). C4A CNVs are predominantly present on the adverse haplotype H2 (carrying the G allele of rs204993) and the protective haplotype H3 (carrying the A allele of rs429608). OR = Odds ratio, [95 % CI] = 95 % confidence intervals, SD = standard deviation. The haplotypes were numbered according to their frequency. (TIF 2488 kb

Additional file 7: Figure S3. of Multiallelic copy number variation in the complement component 4A (C4A) gene is associated with late-stage age-related macular degeneration (AMD)

Sensitivity analysis for protective haplotype H3 at the C2/CFB locus. Phase at C2/CFB was assessed with SHAPEIT2 for each individual. Haplotypes are characterized by the presence or absence of AMD associated alleles from four inpendent variations at this locus (rs429608, rs114190211, rs204993 and rs142511358 [9]). Odds ratios and corresponding 95 % confidence intervals [95 % CI] are given with the size of each rectangle representing the respective relative number of cases and controls for each subgroup. The protective effect of haplotype H3 decreases with age (becomes less protective). (TIF 1225 kb

Mutational analysis of <i>SRY</i> and <i>WT1</i>.

<p>(A) wild type (upper panel, control) and mutated sequence (lower panel, patient) of <i>SRY</i>. (B) schematic representation of the SRY protein. The K128R mutation resides in the HMG domain, just before the cNLS. (C) <i>In vitro</i> luciferase assays of SRY-WT (wild-type) and SRY-K128R (mutant) in HEK293T cell line. Cells were co-transfected with TESCO-E1b-<i>Luc</i>, SF1 and WT or mutant SRY to assess for activation of TESCO. The mean percentages of fold change of luciferase activity of TESCO-E1b-<i>luc</i> over the empty vector, relative to WT SRY levels from six independent assays (each performed in triplicate) are shown. Error bars represent standard error of the mean (SEM). (D) pcDNA3-FLAG-SRY wild-type (WT, 2 µg) or pcDNA3-FLAG-SRY mutant (K128R, 2 µg) were transiently transfected into HEK293T cells using Fugene 6. Exogenous SRY (WT or K128R) expression was detected using a FLAG antibody and a green fluorescent Alexa-488 dye coupled secondary antibody. Nuclei were stained with 4′,6-diamino-2-phenylindole (DAPI). Both wild type and mutant SRY show strong nuclear staining. (E) SRY fluorescence was quantified as previously described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040858#pone.0040858-Argentaro1" target="_blank">[35]</a>. Nuclear accumulation of SRY (WT or K128R) expressed as fluorescence in the nucleus over that in the cytoplasm (Fn/c) were background fluorescence has been subtracted. Measurements represent the average of 3 independent transfections. Results are relative to WT transfected cells (Fn/c given value of 100%). The number of cells analysed is n = 111 (WT) and n = 121 (K128R). Error bars represent the standard error of mean values. Two-tail t-Test of unpaired sample means was performed between WT transfected cells and mutant transfected cells and showed no significant differences. P = 0.49. (F) mutated sequence (upper panel, patient) and wild type sequence (lower panel, control) of <i>WT1</i>, showing the heterozygous +4C&gt;T change.</p

Immunohistochemical staining of the left GB lesion.

<p>(A) representative hematoxylin and eosin (HE) staining. The germ cells present in the GB stain positive for OCT3/4 (B, brown staining), TSPY (C, red staining), and SCF (D, red staining). Supportive cells in the GB stain positive for FOXL2 (E, brown staining), while SOX9 (F) is negative. All slides are counterstained with hematoxylin. Magnification 100x for all.</p