SUPPLEMENTARY MATERIALS

Inflammation Research

HDAC9, TWIST1 and FERD3L gene expression in asymptomatic stable and unstable carotid plaques of atherosclerotic patients.

Silvia Ferronato1*, Matteo Gelati1*, Alberto Scuro2, Silvia Olivato1, Giovanni Malerba1, Maria Grazia Romanelli1, Macarena Gomez Lira1, Carlo Setacci3

1 Department of Neurological, Biomedical and Movement Sciences, University of Verona, Italy

2 Department of Surgery, University of Verona, Italy.

3 Division of Vascular Surgery, University of Siena, Siena, Italy

Corresponding author:

Supplementary Table 1 Characteristics of the Study Patients

Variable / Asymptomatic stable plaques
(ASP) / Asymptomatic unstable plaques
(AUP)
N=6 / N=16
Age, y (mean±sd) / 73±7 / 73±7
Sex, male (%) / 4 (67) / 12 (75)
Risk factors
Hypertension (%) / 5 (83) / 13 (81)
Diabetes (%) / 2 (33) / 0
Smoking (%) / 2 (33) / 5 (31)
History of dyslipidemia (%) / 6 (100) / 10 (63)
NSAID* or glucocorticoid treatment / 0 / 0
Acetylsalicylic acid (%) / 5 (83) / 10 (63)
Statin (%) / 6 (100) / 8 (50)
ACE-inhibitor (%) / 4 (67) / 9 (56)

NSAID*, nonsteroidal antiinflammatory drug

Supplementary Table 2 Primers used in Real Time PCR

Gene / Primer forward (5’-3’) / Primer reverse (5’-3’)
HDAC9 / GCATGAGAACTTGACACGG / TTGTTGCTGTTTTATGGCTAGAAG
TWIST1 / GGCTCAGCTACGCCTTCTC / TGTCCATTTTCTCCTTCTCTGG
FERD3L / GGACACTACGGTGCTGGACT / CCTCCTGGGTCTTCCCTCT
TBP / TGTATCCACAGTGAATCTTGG / ATGATTACCGCAGCAAACC

Supplementary Table 3 Primers and endonuclease enzymes used for genotyping polymorphisms.

Exchanged DNA nucleotides of designed primers are marked as bold characters.

Polymorphism / Primer forward / Primer reverse / Fragment (bp) / Endonuclease
rs2107595 / TGTGGGACAAAAACATTTTCGC / AGGAACTCATCAAATCTGCTAGA / 217 / HhaI

Supplementary Table 4 HDAC9 and TWIST1 gene expression in relation to rs2107595 variant genotypes.

rs2107595
genotypes / HDAC9 gene expression (mean±SD) / TWIST1 gene expression (mean±SD) / P value
GG (n=15)
GA (n=4) + AA (n=3) / 1.17±0.52
1.22±0.73 / 1.10±0.65
1.06±0.49 / Ns
Ns

Ns= not significant

2