2.11 | RT-PCR
According to the manufacturer’s instructions, total RNA was extracted
from LF tissues and cells using the Trizol reagent (Invitrogen) and
converted to cDNA using the Reverse Transcription Synthesis Kit (Takara,
Dalian, China). Then, the gene expression levels were analyzed using a
Thermal Cycler Dice Real-Time system (Takara, China). After the
processes of reaction, the threshold cycle (Ct) was determined and
realtive mRNA level was normalized to the GAPDH expression using the
2-∆Ct method. Designed primers and their sequences
were all summarized in Table 2 and Table 3.