Lightcycler 480 instrument 2 real time pcr system machine

Herein, samples were prepared in the same way as described for proteomics. The RNeasy Mini Kit (QIAGEN, Hilden, Germany) was used to extract RNA based on the manufacturer’s protocols. Then, the genomic DNA was removed, and cDNA was synthesized by HiScript III All-in-one RT SuperMix Perfect for qPCR Kit (Vazyme, Nanjing, China). Moreover, qPCR was carried out using LightCycler 480 SYBR Green I Master (Roche, Basel, Switzerland) on a Roche Diagnostics LightCycler 480 Instrument II Real-time PCR System machine. Subsequently, the glyceraldehyde-3-phosphate dehydrogenase gene was utilized as an internal control to normalize the relative expression levels. The 2^–ΔΔCT method was used to calculate the relative fold changes in the expression level of each gene (40 (link)). Supplementary Table 1 lists all the primers used in the present study.

Herein, samples were prepared in the same way as described for proteomics. The RNeasy Mini Kit (QIAGEN, Hilden, Germany) was used to extract RNA based on the manufacturer’s protocols. Then, the genomic DNA was removed, and cDNA was synthesized by HiScript III All-in-one RT SuperMix Perfect for qPCR Kit (Vazyme, Nanjing, China). Moreover, qPCR was carried out using LightCycler 480 SYBR Green I Master (Roche, Basel, Switzerland) on a Roche Diagnostics LightCycler 480 Instrument II Real-time PCR System machine. Subsequently, the glyceraldehyde-3-phosphate dehydrogenase gene was utilized as an internal control to normalize the relative expression levels. The 2^–ΔΔCT method was used to calculate the relative fold changes in the expression level of each gene (40 (link)). Supplementary Table 1 lists all the primers used in the present study.