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Determination of GPX4 mRNA expression

SH Shuai Huang
BC Bihui Cao
JZ Jinling Zhang
YF Yunfei Feng
LW Lu Wang
XC Xiaopei Chen
HS Hang Su
SL Shengrong Liao
JL Jinggong Liu
JY Jun Yan
BL Baoxia Liang
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After CNE1 cells treatment with CuB (1,10,50 nM) for 24 h, total RNAs were extracted using Trizol reagent (Invitrogen, USA) and dissolved in nuclease-free water. 1 µg RNA was used to reverse transcribe cDNA using Reverse Transcription system Kit (Nanjing Vazyme Biotech Co., Ltd., China) as described in the manufactures’ protocol. cDNA was used for quantitative Real-Time PCR on the QuantStudio 7 Flex (ThermoFisher, MA) using GPX4(fwd: 5′-ACAAGAACGGCTGCGTGGTGAA-3′; rev: 5′-GCCACACACTTGTGGAGCTAGA-3′) or actin (fwd: 5′-ATCTGGCACCACACCTTCTAC-3′; rev: 5′-CAGGTCCAGACGCAGGATG-3′) primers in a SYBR green reaction to determine mRNA levels. GPX4 mRNA expression levels were normalized to the expression of actin mRNA.

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