Quantitative Gene Expression Analysis in Renal Tissue

Gene expression in renal cortical tissue was analyzed by RT-PCR using a LightCycler FastStart DNA Master SYBR Green I kit and an ABI Prism 7000 Sequence Detection System (Applied Biosystems, Foster City, USA) as described previously [18] (link), [23] (link). The oligonucleotide primer sequences for rat β-actin, TGF-β₁, α-SMA, type 1 collagen, MCP-1, PAI-1, gp91phox, Nox-1, E-cadherin, and fibroblast-specific protein 1 (FSP-1) are listed in Table 1. All data are expressed as the relative difference to the 10-week values of the DSS + NS group, after normalization to β-actin expression.

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Rafiq K., Nishiyama A., Konishi Y., Morikawa T., Kitabayashi C., Kohno M., Masaki T., Mori H., Kobori H, & Imanishi M. (2014). Regression of Glomerular and Tubulointerstitial Injuries by Dietary Salt Reduction with Combination Therapy of Angiotensin II Receptor Blocker and Calcium Channel Blocker in Dahl Salt-Sensitive Rats. PLoS ONE, 9(9), e107853.

Publication 2014

LightCycler FastStart DNA Master SYBR Green I kit ABI Prism 7000 Sequence Detection System

Actin Cortical E cadherin Fibroblast specific protein 1 Gene expression Gp91phox Mcp 1 Oligonucleotide primer Pai 1 Prism Renal Rt pcr Sybr green i Tgf β1 Tissue Type 1 collagen

Corresponding Organization :

Other organizations : Kagawa University, Osaka City General Hospital

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Variable analysis

independent variables

Treatment groups: DSS + NS, DSS + RAS inhibitor

dependent variables

Gene expression levels of β-actin, TGF-β1, α-SMA, type 1 collagen, MCP-1, PAI-1, gp91phox, Nox-1, E-cadherin, and fibroblast-specific protein 1 (FSP-1)

control variables

Normalization to β-actin expression

controls

No additional positive or negative controls were explicitly mentioned.

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