Sequencing and Expression Analyses of the Synaptic Lipid Raft Adapter Gene PAG1 In

Supplementary Material

Sequencing and expression analyses of the synaptic lipid raft adapter gene PAG1 in schizophrenia

Shabeesh Balan1, Yoshimi Iwayama1, Kazuo Yamada1, Tomoko Toyota1, Tetsuo Ohnishi1, Manabu Toyoshima1, Chie Shimamoto1,2, Masayuki Ide3, Yasuhide Iwata4, Katsuaki Suzuki4, Mitsuru Kikuchi5, Tasuku Hashimoto6, Nobuhisa Kanahara6, Takeo Yoshikawa1* and Motoko Maekawa1

1Laboratory for Molecular Psychiatry, RIKEN Brain Science Institute, Saitama, Japan;

2Graduate School of Humanities and Sciences, Department of Life Science, Ochanomizu University, Tokyo, Japan;

3Department of Psychiatry, Division of Clinical Medicine, University of Tsukuba, Ibaraki, Japan,

4Department of Psychiatry and Neurology, Hamamatsu University School of Medicine, Shizuoka, Japan;

5Department of Psychiatry and Neurobiology, Kanazawa University Graduate School of Medicine, Kanazawa, Japan;

6Department of Psychiatry, Graduate School of Medicine, Chiba University, Chiba, Japan

*Corresponding author:

Takeo Yoshikawa, MD, PhD

Laboratory for Molecular Psychiatry

RIKEN Brain Science Institute

2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan

Tel: +81(Japan)-48-467-5968

Fax: +81(Japan)-48-467-7462

E-mail:

Brain Region / Diagnosis / Storage time (years)* / Age* / pH* / PMI* / Gender, n (%) / Ethnicity, n (%)
Female / Male / Black / White
BA46 / Control (n= 35) / 16.60 ± 4.49 / 45.85 ± 13.03 / 6.60 ± 0.20 / 13.65 ± 6.36 / 13(0.371) / 22(0.628) / 14(0.400) / 21(0.600)
BA46 / Schizophrenia (n= 35) / 15.68 ± 5.40 / 45.51 ± 13.59 / 6.56 ± 0.44 / 14.20 ± 6.57 / 9(0.257) / 26(0.743) / 11(0.314) / 24(0.686)
CA1 / Control (n=20) / 15.00 ± 4.01 / 42.40 ± 12.30 / 6.60 ± 0.24 / 15.10 ± 6.71 / 7(0.350) / 13(0.650) / 9(0.450) / 11(0.550)
CA1 / Schizophrenia (n=20) / 12.85 ± 5.42 / 45.00 ± 15.34 / 6.37 ± 0.37 / 14.70 ± 5.92 / 5(0.250) / 15(0.750) / 5(0.250) / 15(0.750)

PMI=Post mortem interval, * Mean ± SD

Supplementary Table 1: Clinical characteristics of post-mortem tissue samples from BA46 and CA1 regions, obtained from Maryland Brain Collection ( at the Maryland Psychiatric Research Center, Baltimore, Maryland.

PAG1 coding exons# / Primer Name / Primer sequence / Product Length / Annealing Temperature (oC)
Exon4 / PAG1_E04-1F / GGTCTTGCTCCTGACAGGGA / 410 / 61.0
PAG1_E04-1R / CACAAGTTTCTGGGACGGCT
Exon5 / PAG1_E05-1F / TTGAGGCTTGAACTCCTCTTCC / 403 / 61.0
PAG1_E05-1R / TGCATCCATTTCAGAAGCCA
Exon6 / PAG1_E06-1F / CCACTGGGGGTCTTGAAATG / 476 / 60.5
PAG1_E06-1R / AGTCTTAGAACAATGCCTGGCAC
Exon7 / PAG1_E07-1F / AGCACGCATCTTGATTTAACCTAA / 500 / 60.0
PAG1_E07-1R / CAAACTCAGCTTTGCCTTCAGT
PAG1_E07-2F / GGTGGAGGACTGCTTGTATGA / 494 / 58.5
PAG1_E07-2R / ACACAGCTTACATGAAAACAGCTC
Exon8 / PAG1_E08-1F / TCCCCGAACATCATACATGTG / 460 / 60.0
PAG1_E08-1R / TCCTGAGCTGACTACTTGTCCTG
Exon9 / PAG1_E09-1F / AGGAGTACACACACAGAAACCCAG / 628 / 61.0
PAG1_E09-1R / TGAACAACTGGGAGAACAGTCG

#The PAG1gene comprises9 exons of which6 code for the protein (exons 4 to 9).

Supplementary Table 2: List of primers and conditions used for PAG1 amplification