Quantifying LINC01605 Expression in Healthy and HS Tissues

Total RNA was extracted from HS tissues, healthy tissues and HTFs (5x10⁴/ml) using TRIzol^® reagent (Invitrogen; Thermo Fisher Scientific, Inc.). Total RNA was reverse transcribed into cDNA using the EntiLink™ 1st Strand cDNA Synthesis kit [ELK (Wuhan) Biotechnology Co., Ltd.] according to the manufacturer's instructions. qPCR was subsequently performed using the StepOne™ Real-Time PCR System (Thermo Fisher Scientific, Inc.). EnTurbo™ SYBR Green PCR SuperMix kit (ELK Biotechnology Co., Ltd.) was used for qPCR. The thermocycling conditions were used as follows: 3 min at 95˚C, followed by 40 cycles of 10 sec at 95˚C, 30 sec at 58˚C and 30 sec at 72˚C. mRNA expression levels were normalized to β-actin levels. Relative expression levels were calculated using the 2^-ΔΔCq method (24 (link)). The following primer sequences were used for qPCR: β-actin forward, 5'-GTCCACCGCAAATGCTTCTA-3' and reverse, 5'-TGCTGTCACCTTCACCGTTC-3'; and LINC01605 forward, 5'-CAACTCATTCCCGTTACAAACA-3' and reverse, 5'-CATCTCAACTGCCTCTGTCTCC-3'.