Hek 293t

Pulmonary alveolar macrophages (PAMs) were obtained from lung lavage samples of 4-week-old pigs. CRL-2843-CD163 (the continuous PAM cell line stably expressing porcine CD163), MARC-145 (the derivative from African green monkey kidney cell line MA-104) and HEK-293T (human embryonic kidney 293 cell line stably expressing SV40 large T antigen) cells were used in our studies. HEK-293T and MARC-145 cells were maintained in Dulbecco modified Eagle medium (DMEM, Solarbio life sciences, Beijing, China) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Gibco, Logan, UT, USA) and penicillin–streptomycin mixtures (Solarbio life sciences). PAMs and CRL-2843-CD163 cells were cultured in Roswell Park Memorial Institute-1640 medium (RPMI-1640, Solarbio life sciences) supplemented with 10% FBS and antibiotics.
A typical PRRSV-2 strain BJ-4 (GenBank accession no. AF331831) was a gift from Professor Hanchun Yang of China Agricultural University. rBJ4-EGFP was constructed by inserting enhanced green fluorescent protein (EGFP) between open reading frame (ORF) 1b and ORF2a of PRRSV strain BJ-4 in our laboratory. These viruses used in our study were propagated in MARC-145 cells in DMEM with 3% FBS, and the virus titers were measured by 50% tissue culture infective dose (TCID₅₀) assay in MARC-145 cells [16 (link)].

The LUAD cell lines (H322, SPC‐A‐1, PC‐9, H1299, H1975, A549, H1650, HCC827), NSCLC cell lines (H460, SK‐MES‐1) and HEK293T cells were purchased from American Type Culture Collection (ATCC). Human bronchial epithelial cell (HBEC) was a gift from School of Medicine, Zhejiang University. The LUAD cell lines, NSCLC cell lines and HBEC were maintained in RPMI 1640 medium (Solarbio). HEK293T cells were maintained in DMEM (Solarbio, China). All medium were complemented with 10% foetal bovine serum (FBS, Noverse, 100 units/mL penicillin and 100 μg/mL streptomycin (HyClone). All cells were maintained in a humidified incubator containing 5% CO₂ at 37℃. An AKT inhibitor MK‐2206 was purchased from Meilunbio.