Pgl3 promoter

The plasmid vector pCDH-CMV, pLKO.1-puro, pGL3-Promoter, pRL-CMV, VSVG and GAG-POL were obtained from Addgene (Addgene, USA). The coding sequence of MKL-1 was inserted into the plasmid pCDH-CMV and used to construct the stable overexpression plasmid pCDH-MKL-1 of MKL-1. Plasmid pLKO.1-puro was used to construct MKL-1 knockdown plasmids pLKO.1-sh-MKL-1-1, pLKO.1-sh-MKL-1-2 and pLKO.1-sh-MKL-1 -3. Plasmid pGL3-Promoter was used to construct luciferase reporter plasmids for the SLC3A2 and SLC7A11 promoters. Plasmid pRL-CMV was used as an internal control plasmid for dual-luciferase reporter activity assays. Plasmids VSVG and GAG-POL were used as lentiviral packaging plasmids. The primer sequences and shRNA sequences were entrusted to Shanghai Sangon Bioengineering Co., Ltd. (Sangong, China) to synthesize.

cDNAs of HBx, FoxP3 and E2F1 were amplified and sub-cloned into pcDNA3.1-Flag (Invitrogen, Carlsbad, CA, USA). 3′-UTR fragment of wild-type (WT) CDH2 and mutant (MUT) CDH2 were cloned into the dual-luciferase expression vector pmirGLO (Invitrogen, Carlsbad, CA, USA). The promoter region of WT and MUT of miR-187-5p (WT-miR-187-5p-Luc and MUT-miR-187-5p-Luc), WT and MUT of FoxP3 (WT-FoxP3-Luc and MUT-FoxP3-Luc) were cloned into the luciferase vector pGL3-Promoter (Addgene, Watertown, MA, USA). All cloning primers have been shown in Supplementary Table 1.