Colon Tissue and Caco-2 Cell RNA Extraction and Expression Analysis

RNA was extracted from colon tissue and Caco-2 cells by TRIzol, and reverse transcription (Takara, Japan) and real-time PCR (Vayme, China) were performed according to the instructions of the kit. All primer sequences were synthesized by Sangon Biothen Co., Ltd. (Shanghai, China), and the primer sequences are shown in Table S1. All qRT-PCR analyses were carried out with a 7500 real-time PCR system (Applied Biosystems). The amplification protocol was as follows: 95 °C for 30 s, 40 cycles 95 °C for 10 s, 60 °C for 35 s, 95 °C for 15 s, 60 °C for 60 s, and 95 °C for 15 s. The relative fold change in expression was finally calculated using the 2^−ΔΔCT method. All samples were assayed in triplicate and expression levels of mRNA were normalized using the endogenous control β-actin [35 (link)].