Power sybr premix ex taqtm 2

The RAW264.7 cells were pretreated with AsC (20 μM) 12 h before exposure to ox-LDL for 24 h. Total RNA was extracted using TRIzol (Invitrogen, Carlsbad, CA, United States). The isolated RNA was reverse transcribed into cDNA using the GoScriptTM Reverse Transcription System (Promega). Then, amplification was carried out using real-time RT-PCR with the Power SYBR Premix Ex TaqTM II (TaKaRa Biotechnology, Dalian, China) in an iQ5 Real-time PCR detection system with analysis software (Bio-Rad, Santa Rosa, CA, United States). Primers (Table 1) were designed using premier primer Software 6.0 (Canadian Premier Life Insurance Company, ON, Canada). The 2^-ΔCT method was used to analyze the results according to our previous research [24 (link)].

The HUVECs were incubated with ox-LDL and pretreated with GF1 (16 μM) 24 h before the testing. Total RNA was extracted using TRIzol (Invitrogen, Carlsbad, CA, United States). About 2 μg of total RNA was reversely transcribed using the GoScript^TM Reverse Transcription System (Promega). cDNA was synthesized from the isolated RNA, and cycle time values were obtained using real-time RT-PCR with the Power SYBR Premix Ex Taq^TM II (TaKaRa Biotechnology, Dalian, China) in an iQ5 Real-time PCR detection system and analysis software (Bio-Rad, Santa Rosa, CA, United States) as previously described (Qin et al., 2015 (link)). The cycle number at which transcripts could be detected (CT) was normalized to the cycle number of GAPDH gene detection, and referred to as ΔCT. Primers (Table 1) were designed using premier primer Software 6.0 (Canadian Premier Life Insurance Company, ON, Canada).