Gene Expression Analysis by qRT-PCR

cDNA was synthesized from the total RNA with the RT^{2 (link)} First Strand Kit (Qiagen), Superscript VILO IV (Thermo Fisher), or iSCRIPT cDNA kit (BioRad), always with an additional DNase I treatment step. qRT-PCR assays were performed in technical duplicates in 96- or 384-well plates on QuantStudio 7 (Life Technologies) or Bio-Rad CFX 96 instrument, with RT² SYBR Green (Qiagen), POWER SYBR (Thermo Fisher), iTaq SYBR (BioRad) or TaqMan (Thermo Fisher) expression assays (Table S4). The expression of target genes was normalized by geometric means of endogenous controls (GAPDH, HPRT1, TBP or ACTB, as indicated in Table S2), and presented as dCt values relative to endogenous controls (log2 scale).