Sybr green 1 reagent

Total RNA from aortic tissue and BMDMs was extracted with Trizol (Applygen, Beijing) and reverse transcribed by using a reverse transcription system (Applygen, Beijing). Real-time PCR amplification included Applied Biosystems 7500 fast PCR System (Life Technologies, USA) and SYBR Green I reagent (Tiangen Biotech, Beijing). The cycle threshold (Ct) was determined as the number of PCR cycles required for a given reaction to reach an arbitrary fluorescence value within the linear amplification range. Relative quantification was performed according to the 2^-ΔΔCt method, with GAPDH as a reference. The primers for real-time PCR were in Table 1.

Total RNA was isolated and reverse transcribed. RT-PCR amplification involved using the Applied Biosystems 7500 fast PCR System (Life Technologies, USA) and SYBR Green I reagent (#FP205-02; Tiangen Biotech; Beijing). The cycle threshold (Ct) was determined as the number of PCR cycles required for a given reaction to reach an arbitrary fluorescence value within the linear amplification range. Relative quantification was performed according to the 2^-ΔΔCt method, with GAPDH level as a reference. The forward and reverse PCR primers are in Supplementary Table 1.