Supplemental data

Table e-1.Primers sequences and product sizes of first QMPSF*

Gene / Exon number / 5’-3’ Forwardprimer sequence / 5’-3’ Reverse primer sequence / Product size (bp)
PCBD2 / Exon 2 (NM_032151.4) / AAGCAGCAGGATGGTCGGA / CCTGATTAAAATTGTGGAAGGAGAATT / 95
HPRT1 / Exon 3 (NM_000194.2) / GACTGAACGTCTTGCTCGAG / GCTTTGATGTAATCCAGCAGG / 137
PDGFB / Exon 2 (NM_002608.2) / CTCAGCCTTTTGGTGTCTGCC / TCCATTTACCTCCGGGGTCT / 153
SLC20A2 / Exon 3 (NM_006749.3) / TCTGCACCTCTGTACTCTGTATCTGAT / TACCGATTGCGACCAGTGAG / 176
SLC20A2 / Exon 8 (NM_006749.3 / CGACAGCTACTCGAGCTACTGTAAC / GCAGGAAATGGAACAGGAGGT / 202
PDGFRB / Exon 13 (NM_002609.3) / ATCATCTCTGCACCAGGACGC / GAGAGGCTAAGTGTGTGGGGA / 214
PDGFRB / Exon 3 (NM_002609.3) / TTGCTGTCTCTCCTGTTACTTCTG / GTGAGGTTGGTCAGTGTGAGCA / 241
SLC20A1 / Exon 11 (NM_005415.3) / GTTTCCTTGGTCTGCTTCTCTTC / GAATACCTAAGATGGTCCCAAACAT / 260
SLC20A1 / Exon 2 (NM_005415.3) / GTTCTTTTTATGTTTTGTCCTCTTCG / CCCTCCTGCCCCTTTGCC / 289
PDGFB / Exon 5 (NM_002608.2) / AGGTCGGGAGGGCTTGTTTT / CCAGGTGGTCTTCCAGCGTC / 313
PDGFRB / Exon 21 (NM_002609.3) / TACACCCAGCCACCACTTCC / GCCTTGTACTCGGTGTCTGACTTC / 334

*All forward primers are preceded by the following sequence(5’-3’): 6FAM-CGTTAGATAG

All reverse primers are preceded by the following sequence (5’-3’): GATAGGGTTA

Table e-2

Primers sequences and product sizes of PDGFB QMPSF*

Gene / Exon number / 5’-3’ Forward primer sequence / 5’-3’ Reverse primer sequence / Product size (bp)
PCBD2 / Exon 2 (NM_032151.4) / AAGCAGCAGGATGGTCGGA / CCTGATTAAAATTGTGGAAGGAGAATT / 95
PDGFB / Exon 1 (NM_002608.2) / CTCTGCTGCTACCTGCGTCTG / GGGGCGAAGGTAATGAATGA / 104
PDGFB / Exon 1 alt (NM_033016.2) / GACTGAGCAGGAATGGTGAGAT / ATAGGAAGAGTTGTCACAGAAGAGG / 129
PDGFB / Exon 2 (NM_002608.2) / CTCAGCCTTTTGGTGTCTGCC / TCCATTTACCTCCGGGGTCT / 153
PDGFB / Exon 3 (NM_002608.2) / GAGTGGGGGAGAGGCAAAC / AGAGTGGGAGCGGGTCATG / 170
PDGFB / Exon 4 (NM_002608.2) / CGCACCAACGCCAACTTC / GGTCTCCACCCACCACCG / 192
HMBS / Exon 6 (NM_000190.3) / GAAGGCTGGCTGCTCATACC / AAGGCAAAGGTTCACATGATGC / 211
PDGFB / Exon 5 (NM_002608.2) / AGGTCGGGAGGGCTTGTTTT / GACAGAGCCACAAAATGCCTCT / 234
PDGFB / Exon 6 (NM_002608.2) / AGCCAAAACGCCCCAAAC / CCCCTGACCCCATCCACA / 250

*All forward primers are preceded by the following sequence(5’-3’): 6FAM-CGTTAGATAG

All reverse primers are preceded by the following sequence (5’-3’): GATAGGGTTA

Table e-3. Primers used for cDNAPCR amplification, designed to amplify both deleted and wild type allele ofPDGFB(NM_002608.2).

5’-3’ sequence / Forward / reverse / Location
AGGCCTGAGCGCCTGAT / Forward / 5’UTR (untranslated exon 1)
AATGGTCACCCGAGTTTGG / Reverse / Exon 6

Supplementary methods

Skin biopsy was taken from the right buttock of the patient. Punch biopsy was performed, and then fixed in glutaraldehyde solution. After postfixation in buffered osmium tetroxide and dehydratation in ascending grade of ethanol, the samples were embedded in epoxy resin (Epon). We reviewed semi-thin sections stained with toluidine blue to select vessels. Once appropriate vessels located, ultra-thin sections (80 nm) were cut and stained with 2% uranyl acetate and lead citrate and observed in a transmission JEM100-11 electron microscope (Jeol) at the EM laboratory of the Caen University (CEMAbio). Electron images were captured using a CCD camera (GATAN Orius 200) and digitally enregistered with SIS megaview III-logiciel Analysis 5.

Every vascular wall (from artery, vein or capillary), previously located on semi-thin sections, was precisely analyzed. A total of 6 arterioles, 18veinules and 4 capillaries were examined.

Neither granular osmiophilic material nor calcifications were observed in the limit of the skin biopsy.