RNA Isolation, RT-PCR, and Quantification

RNA isolation and PCR amplification were performed as previously described^{38 (link)}, except that the RT–PCR was performed using the Superscript III First-Strand system (Life Technologies). Total RNA was isolated from cell lines with the RNeasy kit (Qiagen). One microgram of purified RNA was reverse transcribed using Superscript III First-Strand (Invitrogen) according to the manufacturer’s protocol, and quantitative PCR was performed using SYBR Green on a Viia7 Real-Time PCR system (Thermo Fisher Scientific). All experiments were performed in biological triplicates unless stated otherwise. Each individual biological sample was amplified by qPCR in technical replicates and normalized to actin as an internal control. Amplification was carried out with primers specific to the genes to be quantified (sequences available on request).