Quantifying Gene Expression in Venous Thrombi

Total RNA was extracted from the venous thrombi using the TRIzol reagent (Invitrogen). The cDNA was prepared from DNase-treated RNA (Applied Biosystems). Real-time PCR was performed with ABI PRISM 7900 using TaqMan Universal PCR Master Mix and TaqMan Gene Expression Assays (Applied Biosystems) [17 (link)]. The following TaqMan Gene Expression Assays were used for each gene: assay ID Rn01538170_m1 for MMP-2, assay ID Rn00579162_m1 for MMP-9, assay ID Rn00565261_m1 for urokinase-type plasminogen activator (uPA), assay ID Rn01482578_m1 for tissue-type plasminogen activator (tPA), assay ID Rn01481341_m1 for plasminogen activator inhibitor-1 (PAI-1), assay ID Rn1462586_m1 for fibrinogen α chain, and assay ID Rn99999916_s1 for glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH was used as an internal control.

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Oboshi M., Naito Y., Sawada H., Hirotani S., Iwasaku T., Okuhara Y., Morisawa D., Eguchi A., Nishimura K., Fujii K., Mano T., Ishihara M, & Masuyama T. (2015). Temporary Dietary Iron Restriction Affects the Process of Thrombus Resolution in a Rat Model of Deep Vein Thrombosis. PLoS ONE, 10(5), e0126611.

Publication 2015

TRIzol reagent DNase-treated RNA PRISM 7900 TaqMan Universal PCR Master Mix TaqMan Gene Expression Assays

Assay Cdna Dnase Fibrinogen Gene Gene expression Glyceraldehyde 3 phosphate dehydrogenase Mmp 2 Mmp 9 Plasminogen activator inhibitor 1 Prism Real time pcr Thrombi Tissue type plasminogen activator Trizol Urokinase type plasminogen activator Venous

Corresponding Organization : Hyogo Medical University

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Variable analysis

independent variables

None explicitly mentioned.

dependent variables

MMP-2 expression
MMP-9 expression
Urokinase-type plasminogen activator (uPA) expression
Tissue-type plasminogen activator (tPA) expression
Plasminogen activator inhibitor-1 (PAI-1) expression
Fibrinogen α chain expression

control variables

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression was used as an internal control.

controls

No positive or negative controls were explicitly mentioned.

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