C1000 real time cycler

Total RNA was extracted using Trizol reagent (Invitrogen, Life Technologies). The cDNA was synthesized from 3 μg of total RNA using the Reverse Transcription System (Promega, Madison, WI, USA) according to the manufacturer’s instructions. All qRT-PCR reactions were performed using the C1000 Real-Time Cycler (Bio-Rad Laboratories, Hercules, CA, USA) and qRT-PCR Master mixes (Promega, Madison, WI, USA). Primers for amplification of the MMP7, MMP9, p65, p50 and ACTIN genes are listed in Additional file 1: Table S1. Expression of target genes was determined according to the 2^-ΔΔt method using ACTIN as a reference gene. All experiments were carried out in triplicate.

The RNA of cells was isolated using the Trizol reagent (Invitrogen, Life Technologies), and conversion of total RNA to complementary DNA was performed with the Reverse Transcription System (Promega, Madison, WI, USA). RT-PCR was performed using the Master mixes (Promega, Madison, WI, USA) following the manufacturer’s instructions. All qRT-PCR reactions were performed using the C1000 Real-Time Cycler (Bio-Rad Laboratories, Hercules, CA, USA) and qRT-PCR Master mixes (Promega, Madison, WI, USA). Primers for amplification of the TC2N, p53, p21, BAX, Bcl-2 and ACTIN genes are listed in Supplementary Table S3. All experiments were carried out in triplicate, and the 2^‒ΔΔct method was used to determine expression of the genes of interest.

The RNA of cells was isolated using the Trizol reagent (Invitrogen, Life Technologies), and conversion of total RNA to cDNA was performed with the Reverse Transcription System (Promega, Madison, WI, USA). All qRT-PCR reactions was performed using the C1000 Real-Time Cycler (Bio-Rad Laboratories, Hercules, CA, USA) and qRT-PCR Master mixes (Promega, Madison, WI, USA). Primers for amplification of the DSC1, DSG1, DSC2, JUP, DSP, PKP1, DSC3, DSG3, PKP3 and ACTIN genes are listed in Additional file 1: Table S1. All experiments were carried out in triplicate, and the 2^-ΔΔt method was used to determine expression of the genes of interest.