Invitrogen trizol reagent kit

Total RNA was isolated from cell samples using an Invitrogen™ TRIzol^® reagent kit (Thermo Fisher Scientific, Inc., Waltham, MA, USA), following the manufacturer’s protocol. RNA was reverse-transcribed into cDNA using a PrimeScript™ RT Reagent Kit (Takara Bio, Inc., Kusatsu, Japan) followed by qPCR analysis using SYBR™ Green PCR Master mix (Promega Corporation, Madison, WI, USA) on a CFX96 PCR Detection System. The sequences of the primers used in the present study were as follows: E-cadherin, forward, 5′-GGCCCAGGAGCTGACAAAC, and reverse, 5′-CC AGAGGCTGCGTCACTTTC; vimentin, forward, 5′-ATGAAAGTGTGGCTGCC AAGAAC, and reverse, 5′-GTG ACTGCACCTGTCTCCGGTA; and α-SMA, forward, 5′-CCTCTTCCAGCCATCTTTCAT, and reverse, 5′-CGAGAGGACGTTGTTAGCAT AG. Quantification of the cDNA was determined using the 2^−ΔΔCq method.

RNA was extracted from collected samples of 12-2 using the Invitrogen TRIzol Reagent kit (Thermo Fisher Scientific, Shanghai, China). The concentration and integrity of total RNA were determined by 0.8% agarose gel electrophoresis and a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Worcester, MA, United States). cDNA was synthesized by reverse transcription using the One-Step gDNA removal and cDNA synthesis kit (Transgen Biotech, Beijing, China).