Quantitative analysis of CTSL expression

RNAs were extracted from OV2008 and C13 cells treated with QC at 0, 5 and 10 µM using Qiagen RNA isolation kits following manufacturer’s instruction. About 1 μg of RNA was reverse transcribed using the QuantiTect Reverse Transcription cDNA synthesis kit (Qiagen, Germantown, MD, USA). Quantitative real-time PCR (qRT-PCR) was carried out using SYBR-Green PCR Master Mix (Applied Biosystems, Foster City, CA, USA) in CFX96 Real-Time PCR System (Bio-Rad, Hercules, CA, USA) as reported previously [18 (link)] by using primers synthesized from Integrated DNA Technologies (IDT). The sequences for the genes analyzed are CTSL FP:5′-CAATCAGGAATACAGGGAAGGG-3′, CTSL RP:5′-CTGGGCTTACGGTTTTGAAAG-3′, and GAPDH FP:5′-ACATCGCTCAGACACCATG-3′ and GAPDH RP:5′-TGTAGTTGAGGTCAATGAAGGG-3′.