The mRNA expression in renal cortical tissues was analysed by RT-PCR using a LightCycler FastStart DNA Master SYBR Green I kit and an ABI Prism 7, 000 Sequence Detection System (Applied Biosystems, Foster City, USA) as previously described [15 (link), 35 (link)]. The oligonucleotide primer sequences for rat and human are listed in ESM Tables 1 and 2 , respectively. All data from in vivo studies are expressed as the relative difference in expression compared with LETO rats after normalisation for β-actin expression. Data from in vitro studies are expressed as the relative difference in expression compared with 5 mmol/l glucose after normalisation for β-actin expression.
Protocol detail
Renal mRNA Expression Analysis
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Actin
Glucose
Human
Mrna
Oligonucleotide primer
Prism
Rats
Renal cortical
Rt pcr
Sybr green i
Tissues
Corresponding Organization :
Other organizations : Kagawa University, University of Würzburg, Ehime University
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Variable analysis
independent variables
- MRNA expression in renal cortical tissues
- Relative difference in expression compared with LETO rats after normalisation for β-actin expression
- Relative difference in expression compared with 5 mmol/l glucose after normalisation for β-actin expression
- LETO rats
- 5 mmol/l glucose
- Positive control: LETO rats
- Negative control: 5 mmol/l glucose
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