Huh7 cells

HEK293, HepG2 and Hep3B cells were obtained from the American Type Culture Collection (ATCC), while BEL7402 and SMMC7721 cells were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Huh7 cells were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China). All cells were cultured in RPMI-1640 containing 10% FBS (Gibco, life technologies, California, USA) and penicillin/streptomycin (Solarbio, beijing China) at 37 ℃ in a 5% CO₂ incubator.
In the indicated experiments, cells were transfected with control, TRAIL, or IER3 overexpression vectors using plasmids prepared by GeneChem (Shanghai, China). Briefly, cells were plated in 6-well plates and grown overnight until 70–80% confluent, at which time Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, MA, USA) was used to transfect cells. At 8 h post-transfection, media was replaced and cells were cultured for an appropriate time prior to downstream experimentation.
The pAdEasy-1 vector in E. coli BJ5183 was used to prepare recombinant adenoviruses, as in prior studies [24 (link)] An appropriate multiplicity of infection was determined by comparing total numbers of infected cells to total plaque-forming units, with duplicate samples being used to titrate prepared adenoviruses to titers of 1 × 10¹¹ pfu/ml.

THLE-3 cells were purchased from the American Type Culture Collection (Rockville, MD, USA). Huh7 cells were obtained from Procell Life Science&Technology Co., Ltd. (Wuhan, China). The Hep-3B, HCCLM3 and MHCC97H cell lines were purchased from Beyotime Biotechnology (Shanghai, China). THLE-3 and Hep-3B cells were cultured in MEM (#E600020), while Huh7, HCCLM3, and MHCC97H cells were cultured in DMEM (#E600004). All media were replenished with 10% fetal bovine serum (FBS, #E600001) and 1 × penicillin–streptomycin solution (#E607011) before use, and were refreshed every 3 days. Cells were cultured under 5% CO₂ at 37°C. The reagents used in the cell culture were obtained from the BBI Life Sciences Corporation (Shanghai, China).