The total RNA was extracted from kidney tissues using Trizol reagent (Invitrogen, Carlsbad, CA, USA). The mRNA levels of podocin and nephrin were determined by quantitative RT-PCR. The primers used were as follows: podocin, 5′-CTTGGCACATCGATCCCTCA-3′ and 5′-CTCTCCACTTTGATGCCCCA-3′; nephrin, 5′-AACCGAGCCAAGTTCTCCTG-3′ and 5′-GGACGACAAGACGAACCAGT-3′; GAPDH, 5′-GGAGTCTACTGGCGTCTTCAC-3′ and 5′-ATGAGCCCTTCCACGATGC-3′.
Protocol detail
Quantification of Podocin and Nephrin mRNA
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Corresponding Organization :
Other organizations : Yangpu Hospital of Tongji University, Shanghai University of Traditional Chinese Medicine, YangPu Geriatric Hospital
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Variable analysis
dependent variables
- MRNA levels of podocin
- MRNA levels of nephrin
- RNA extraction method (Trizol reagent)
- Quantitative RT-PCR method
- Primer sequences for podocin, nephrin, and GAPDH
- No positive or negative controls were explicitly mentioned.
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