Quantitative Real-Time PCR

Total RNA was extracted from kidney tissue using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Then, 1 µg RNA was reverse transcribed into complementary DNA (cDNA) using HiScript II Q RT SuperMix for qPCR (Vazyme, NJ, China) in accordance with the manufacturer’s instructions. Real-time PCR was performed as described in our previous report using a Mastercycle reprealplex system (Eppendorf, Hamburg, Germany). Target gene expression levels were evaluated by reference to the value of GAPDH using the 2^−ΔΔCt method. The PCR primers (including those for fibronectin, collagen I, E-cadherin, α-SMA, TGF-β1, Snail, and GAPDH) were synthesized according to sequences provided by PrimerBank ( Table 1 ).

Primers sequences used for RT-PCR.