Mission short hairpin rna shrna lentiviral transduction particles

The human normal liver cell line named MIHA and the Hep3B hepatoma cell line were purchased from American Type Cell Culture (ATCC). The metastatic HCC cell line MHCC97L (97L in short) provided by the Liver Cancer Institute & Zhongshan Hospital of Fudan University, Shanghai, People's of Republic of China (27) . The cell lines were cultured in DMEM high glucose (Gibco; Thermo Fisher Scienti c, Inc.) with 10% FBS (Gibco; Thermo Fisher Scienti c, Inc.), 1% penicillium and streptomycin in a 37 ˚C incubator supplied with 5% CO 2 .
The lentiviral open reading frame (ORF) clone of monomeric green uorescent protein (GFP)-tagged human GSTA2 (cat. no. RC202479L2) and lentiviral ORF control clone were purchased from OriGene Technologies, Inc. Lentiviral particles were produced by transfection into HEK293 cells using Lenti-vpak Lentiviral Packaging kit (OriGene Technologies, Inc). Suppression of GSTA2 was conducted by GSTA2-speci c Mission® short hairpin RNA (shRNA) lentiviral transduction particles (Sigma-Aldrich; Merck KGaA). Mission TurboGFP Control transduction particles (Sigma-Aldrich; Merck KGaA) was used as a control. Lentiviral particles were transduced into HCC cells according to the manufacturer's instructions. The expression levels of GSTA2 mRNA and protein was examined by RT-PCR) and Western blot respectively.