Total RNA was extracted from cells using TRIzol reagent (Life Technologies). Purity and integrity of the RNA were confirmed spectroscopically and by gel electrophoresis before use. One microgram of total RNA was reverse transcribed in a final volume of 20 μL using the RETROscript kit (Life Technologies) and cDNA was diluted 1:3 in nuclease-free water. The evaluation of TFF1, CTSD, CCND1 and MYC mRNA expression was performed by real-time RT-PCR, using SYBR Green Universal PCR Master Mix (Bio-rad). The relative gene expression levels were calculated using the ΔΔCt method as described (Catalano et al., 2015 (link)). Primers are listed in Supplementary Table 2 .
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