RBE and HCCC9810 cell lines were obtained from the Cell Bank of Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences. HuH28, HuCCT1, and CCLP1 were provided by Prof. Qiang Gao (Liver Cancer Institute, Zhongshan Hospital, Fudan University). RBE and HCCC9810 were cultured in Dulbecco's modified Eagle's medium (Gibco, 31885–023) and CCLP1 were cultured in 1,640 medium (Gibco, 72400047) supplemented with 10% fetal bovine serum (Gibco, 10099–141), 20 U/mL penicillin and 20 μg/mL streptomycin (Sigma-Aldrich), with a 5% CO2 concentration at 37°C. HuH28 was cultured in MEM medium (Gibco, 42360032) supplemented with 15% fetal bovine serum (Gibco, 10099–141), 20 U/mL penicillin and 20 μg/mL streptomycin (Sigma-Aldrich), with a 5% CO2 concentration at 37°C.
Full-length cDNA encoding human KDM5C and FASN were cloned into pCDH-CMV-MCS-EF1-Puro (CD510B-1, System Biosciences) with or without Flag-tagged using standard protocols. The shRNA sequences for KDM5C and FASN were purchased from Sigma-Aldrich and cloned into pLKO.1 TRC (Addgene plasmid 10,879). Details about the sequences used here can be found inTable S3 . We used a scrambled siRNA precursor (Scr) as control. DNA sequencing and western blot were performed to verify the constructions. Plasmids and shRNAs were transfected into cell lines through lipofectamine 2,000 (Invitrogen) like previous descriptions (32 (link)).
Full-length cDNA encoding human KDM5C and FASN were cloned into pCDH-CMV-MCS-EF1-Puro (CD510B-1, System Biosciences) with or without Flag-tagged using standard protocols. The shRNA sequences for KDM5C and FASN were purchased from Sigma-Aldrich and cloned into pLKO.1 TRC (Addgene plasmid 10,879). Details about the sequences used here can be found in