Talent qpcr kit

The cells were collected and washed by 4°C PBS. Then, the RNeasy Mini kit (Qiagen, Germany) was used to extract the total RNA and reversed by ThermoScript RT-PCR System (Invitrogen, Carlsbad, CA, United States). The mRNA expression was detected by Talent qPCR kit (TIANGEN, Beijing, China). The primers used in this study are shown in Table 1.

Total RNA of renal tissues was extracted with TRIzol reagent (DP405-02, Tiangen Biotech Co., Ltd., Beijing, China) according to the manufacturer’s standard protocols. cDNA was synthesized using the First Strand cDNA Synthesis Kit (KR118, Tiangen Biotech Co., Ltd., Beijing, China). Amplification reactions were administrated using Talent qPCR kit (FP209, Tiangen Biotech Co., Ltd., Beijing, China). qPCRs were run in a 3,000 Fast Real-Time PCR System (Stratagene Mx3000P, Agilent Technologies Inc., Santa Clara, CA, United States). The 2^–ΔΔCt methods were used for relative mRNA quantification with β-actin as an internal control. Three replicates for each set of samples. Primer sequences are listed in Table 1.

The drug-treated cells were collected and washed by PBS, and then using the RNeasy Mini Kit (Qiagen, Germany) the total RNA is extracted and reversed to cDNA by Bio-Rad RT-PCR System (Hercules, CA, USA). Follow the instructions of the Talent qPCR kit (TIANGEN, Beijing, China) to configure the reaction mixtures, and then incubate it at 95°C for 5 min, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min using the ABI 7500 Fast System (Applied Biosystems, New York, USA). Data were normalized to GAPDH. The primers used in this study are shown in Table 1.