Expression and biochemical results.

<p>A) CNP plasma concentration in patient 2, his parents and three controls. The concentration of the peptide was increased more than 4-fold in the propositus compared to parents and unrelated controls (n = 3). B) Analysis of NPPC expression by real-time PCR analysis in lymphoblasts from patient and his parents. The expression of NPPC in the propositus was remarkably higher compared to those of his father and mother.</p

Clinical signs of the patient 1.

<p>The patient 1 at 3 years of age: short stature, prominent forehead, strabismus, epicanthus, low-set-ears, depressed nasal bridge, short phyltrum, V-shaped mouth. Mild brachydactyly, cutaneous syndactyly of 2nd and 3rd toes.</p

Clinical signs of the patient 2.

<p>A) Details of feet, showing very long hallux bilaterally B) X-rays, showing severe scoliosis. C) Long and curved lower limbs.</p

Expression and biochemical results.

<p>A) CNP plasma concentration in patient 1, his parents and six controls. The concentration of the peptide was normal compared to parents and unrelated controls (n = 6). B) Analysis of <i>NPPC</i> expression by real-time PCR analysis in lymphoblasts from patient, his parents, and ten controls. The expression of NPPC in the proposita was normal compared to those of his parents and unrelated controls (n = 10).</p

Schematic representation of the genomic region spanning the described deletions according to the UCSC Genome Browser Feb 2009 (GRCh37/hg19).

<p>Several annotated genes and ESTs are present, the two deletions are indicated by black arrows (patient 1 and patient 2), whereas triangles indicate the position of the translocation breakpoints previously published <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066048#pone.0066048-Bocciardi1" target="_blank">[11]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066048#pone.0066048-Moncla1" target="_blank">[12]</a>.</p

Array-CGH and FISH results.

<p>A) Result of array-CGH analysis of chromosome 2 with Agilent Human Genome CGH microarray Kit G3 400 in patient 1. The 1.914 Mb interstitial q37.1 deleted region of chromosome 2 extends between oligomers A_18_P13670199 (231,264,956 bp) and A_16_P00618306 (233,178,325 bp) flanked by oligomers A_16_P00615757 (231,257,468 bp) and A_16_P00618312 (233,181,399 bp) (UCSC Genome Browser, <a href="http://genome.ucsc.edu/" target="_blank">http://genome.ucsc.edu/</a>; February 2009). B) FISH with BAC clones RP11-395A23 (AC010149) (chr2:231,304,236–231,476,367). The arrowhead indicates the deleted chromosome 2. D) Result of array-CGH analysis of chromosome 2 with Agilent Human Genome CGH microarray Kit G3 400 in patient 2. The 4.515 Mb interstitial deletion at bands q37.1q37.3 of chromosome 2 was comprised between oligomers A_16_P16076619 (232,963,736 bp) and A_16_P36124457(237,479,062 bp) flanked by oligomers A_16_P16076610 (232,954,321 bp) and A_16_P36124475 (237,483,914 bp). C) FISH with RP11-485M18 (AC079400)(chr2:236,766,818-236,919,215). The arrowhead shows the deleted chromosome 2.</p

Additional file 1: of Assessment of copy number variations in 120 patients with Poland syndrome

Figure S1. Quantitative polymerase chain reaction. In the Figure, examples of qPCR results obtained from three PS patients carrying three different CNVs are shown. A) Patient PS16: primers were designed to amplify a region encompassed by the duplication (chr16:74,313,856-74,313,966) and a 5′ flanking region (chr16:73,857,457-73,857,556); B) patient PS3: primers were designed to amplify a region encompassed by the duplication (chr5:22,539,039-22,539,170) and a 5′ flanking region (chr5:21,756,984-21,757,122); C) Patient PS10: primers were designed to amplify a region encompassed by the deletion (chr11:28,126,119-28,126,268) and a 5′ flanking region (chr11:28,075,866-28,076,003). Fold change of about 1 is expected for a diploid sample, about 0.5 for a haploid sample, and about 2.0 for a triploid sample. ctr: genomic DNA from one healthy adult used as control. (TIF 1168 kb