2.8 qPCR
Total RNA of colonic tissues and cells were extracted by using Trizol
(Accurate Biotechnology, Hubei, China) accroding to manufacturer’s
instructions. The cDNA was synthesized by using High Capacity cDNA
Reverse Transcription Kit (ThermoFisher Scientific, NY, USA) and
amplified with SYBR Green Master Mix
(Thermo Fisher Scientific, NY,
Massachusetts, USA). The folding changes of mRNA expression of target
gene were normalized to GAPDH by ΔΔCt method. The unit
of the variable is determined as the folded mean of the control.