Fig. 2

Characterization of the duplications. a–c Characterization of the 0.88 Mb duplication in family 1. a Analysis of 11p15 microsatellite markers showing the segregation of the duplication in three generations. The haplotype of the chromosome carrying the duplication is shadowed. Gray color of II-2 and III-2 indicates a growth restriction observed only in childhood. b DNA methylation analysis of ICR1 and ICR2 as determined by Pyrosequencing. Line chart reporting the methylation level (%) of seven CpGs of ICR2 and three CpGs of ICR1.The placenta DNAs of the proband (III-3) and a healthy control (Plac. N 1) and leukocyte DNAs of the parents (II-1 and II-2) and a healthy control (normal Ctrl 1) showed similar methylation patterns at both ICRs. Two BWS patients carrying a duplication of the entire domain [17, 18], Dupl Ctrl, have been analyzed as controls. c Real-time messenger RNA (mRNA) expression analysis of CDKN1C and PHLDA2 normalized to the GAPDH control gene in the placenta cells of the proband (III-3) and three normal controls (Ctrl 1, 2, 3). Experiments were performed in triplicate and statistical significance determined by Student’s t test. d–e Characterization of the 1.13 Mb duplication in family 2. d DNA methylation analysis at ICR1 and ICR2 determined by Pyrosequencing in the trio as described in b. The proband (III-1) and his mother (II-2) show ICR2 methylation level similar to two BWS patients carrying ICR2 duplications (Dupl Ctrl 1 and 2) and lower than the father (II-1) and two healthy controls. ICR1 methylation of the proband and his parents is comparable to that of two healthy controls (normal Ctrl 1 and 2). e Analysis of 11p15 microsatellite markers showing a de novo paternal duplication in the mother and a maternally inherited duplication in the proband. The haplotype of the chromosome carrying the duplication is shadowed