Validation of DEGs by quantitative polymerase chain reaction
(qPCR)
cDNA preparation was achieved using the PrimeScript 1st strand cDNA
Synthesis Kit (Takara Bio). qPCR was implemented using the SYBR
Green Quantitative RT-PCR (Sigma-Aldrich) on an Azure Cielo 3 Real-Time
qPCR System (Azure Biosystems). The PCR cycle condition was 50°C for 2
minutes, 95°C for 10 minutes, 95°C for 15 seconds and 60°C for 30
seconds in 40 cycles, then 72°C for10 minutes. After normalization to
β-actin mRNA level, the relative transcript amounts were determined by
the 2-ΔΔCt method. The primer sequences are displayed
in Supplementary Table 1&2 .