Ag reverse transcription kit

In this study, we use TRIzol reagent (Invitrogen, Carlsbad, CA, United States) to extract total RNA from human nucleus pulposus cells and rat nucleus pulposus tissues.
Total RNA was extracted from human nucleus pulposus cells or rat nucleus pulposus tissues by using the TRIzol RNA isolation protocol (Invitrogen, Inc., Carlsbad, CA, United States). AG Reverse Transcription Kit (Accurate biotechnology, Changsha, Hunan, China) was used to reverse transcribe total RNA into cDNA and SYBR Premix ExTaq II (Yeasen Biotechnology, Shanghai, China) SYBR was used for the reactions of qPCR. Then we used a ABI 7500 sequencing detection system (Applied Biosystems, Foster City, CA, United States) to measure the reactions and applied the β-actin as a control RNA expression level. Data were analyzed by GraphPad Prism (GraphPad Prism Software 6.0, United States). The primers we used in the experiments: β-actin, AGA​GCT​ACG​AGC​TGC​CTG​AC, PLK1, CAC​CAG​CAC​GTC​GTA​GGA​TT, MMP3, CCT​ACA​AGG​AGG​CAG​GCA​AG, MMP13, TCG​GCC​ACT​CCT​TAG​GTC​TT, COL2A1, CCA​GAT​GAC​CTT​CCT​ACG​CC, ADAMTS4, AAC​GTC​AAG​GCT​CCT​CTT​GG, ADAMTS5, CCG​GAG​CCA​CTG​CTT​CTA​TC, aggrecan, GGG​ACC​TGC​AAG​GAG​ACA​GAG, SOX9, GCT​CTG​GAG​ACT​TCT​GAA​CGA, p53, CCA​GGA​TGT​TGC​AGA​GTT​GTT​A, p21, TAT​TTT​GTC​CTT​GGG​CTG​CCT, p16, CTT​CGG​CTG​ACT​GGC​TGG.