Quantitative Assessment of Fer1L4 lncRNA

To determine the lncRNA expression profile of Fer1L4, we performed quantitative real-time PCR (qRT-PCR). Each qPCR was conducted with 5 ng/µl cDNA template and 10 pmol/µl of each forward and reverse primer using SYBR^® Premix Ex Taq™ II with ROX Plus (Takara Bio, Saint-Germain-en-Laye, France). PCR experiments were performed on an ABIPrism 7900 HT Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The following primer sequences were used: Fer1L4 (forward ACA-CAG-TCC-TTG-TGG-GTT-CC; reverse CCT-GTC-TCC-TCC-ATC-TCT-CC). Obtained data were analyzed using Qbase + software (Biogazelle, Ghent, Belgium) with ACTB and PPIA as reference genes in the $2^{-\mathrm{\Delta }\mathrm{\Delta }{C}_{\text{T}}}$ algorithm. Both genes were shown to be suitable reference genes for RCC studies [19 (link), 20 (link)].