Verification of ETS1 by RT-qPCR

Ten pairs of clinical ccRCC samples and adjacent kidney tissues frozen in liquid nitrogen were obtained from primary ccRCC patients undergoing radical nephrectomy at the Department of Urology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine. According to the manufacturer’s instructions, total RNA was extracted using TRIzol reagent (Invitrogen, Waltham, MA, USA), cDNA was synthesized and the qRT-PCR was performed and calculated by means of 2^-ΔΔCt methods. GAPDH served as an internal standard control for mRNA expression. Related primers were as follows: ETS1, Gene ID: {"type":"entrez-nucleotide","attrs":{"text":"NM_005238.4","term_id":"1676319366","term_text":"NM_005238.4"}}NM_005238.4, F: 5’-GAAGAGTGGTGGGTGGTTTAT-3’, R: 5’-CAGATTTGCCCATCCTTCCT-3’; GAPDH, Gene ID: {"type":"entrez-nucleotide","attrs":{"text":"NM_001256799.3","term_id":"1676318038","term_text":"NM_001256799.3"}}NM_001256799.3, F: 5’-CAAGAGCACAAGAGGAAGAGAG-3’, R: 5’-CTACATGGCAACTGTGAGGAG-3’. This study was approved by the Institutional Research Ethics Committees of The First Affiliated Hospital of Guangzhou University of Chinese Medicine.