Total RNA extraction and RT-qPCR analysis
Total RNA was extracted using the TRIzol Reagent (CoWin Biosciences,
Jiangsu, China) according to the manufacturer’s instructions. The
extracted RNA was reverse-transcribed into cDNA using Super Script III
Reverse Transcriptase (Vazyme Biotech Co., Ltd., Nanjing, China). ABCB1
mRNA expression was evaluated through a relative quantitative method.
The formula, 2-△△CT, was used to calculate the
quantity of DNA amplified. The sequencing primers used for
amplification, which were provided by Tsingke Biotechnology Co., Ltd.,
were;
ABCB1-F: 5′-TTGCTGCTTACATTCAGGTTTCA-3′,
ABCB1-R: 5′-AGCCTATCTCCTGTCGCATTA-3′,
GAPDH-F: 5′-GCACCGTCAAGGCTGAGAAC-3′,
GAPDH-R: 5′-TGGTGAAGACGCCAGTGGA-3′.