Quantification of lncRNA PART1, SOCS6, and miR-17-5p

Trizol (Invitrogen, USA) was used for segregating total RNA followed by reverse transcription of lncRNA PART1 and SOCS6 with BeyoRT™II Kit (Beyotime, Shanghai, China). Reverse transcription of miR-17a-5p was conducted using Mir-X Kit (Takara, Japan). Then, 7300Plus Real-Time PCR System (Applied Biosystems, USA) with SYBR Green (Applied Biosystem) was applied for RT-qPCR: predenaturation, 95 °C, 30 s and (denaturation, 95 °C, 5 s, annealing, 60 °C, 10 s and extension, 72 °C, 30 s) × 35 cycles. Primers were listed as below, which were: lncRNA PART1 5′-CAATAAGGCAGAAGAAGGTG-3′ (forward), 5′-GGAGAATCTGAAGTCCCAAG-3′ (reverse)[18 (link)]; miR-17-5p 5′-CGGCGGCAAAGTGCTTACAG-3′ (forward), 5′-GTGCAGGGTCCGAGGT-3′ (reverse)[19 (link)]; SOCS6 5′-GGAATTCATGAAGAAAATCAGTCTGAA-3′ (forward), 5′-CGGAATTCTCAGTAGTGCTTCTCCTGCA-3′ (reverse)[20 (link)]; GAPDH 5′-CCTGCCTCTACTGGCGCTGC-3′ (forward), 5′-GCAGTGGGGACACGGAAGGC-3′ (reverse) and U6 5′-CTCGCTTCGGCAGCACA-3′ (forward), 5′-AACGCTTCACGAATTTGCGT-3′ (reverse). GAPDH and U6 were used for normalization of lncRNA PART1, SOCS6, and miR-17-5p, respectively and the relative expression levels were analyzed using 2^-ΔΔCt method.