Huvec cells

HUVEC cells (ScienCell Research Laboratories, Carlsbad, CA, USA) were grown in ECM BulletKit (ScienCell Research Laboratories, Carlsbad, CA, USA) in a 5% CO₂ incubator. A549 cells (ATCC Cat# CRM-CCL-185, RRID:CVCL_0023) were grown in our laboratory in DMEM with 10% FBS (Thermo Scientific, Waltham, MA, USA) in a 5% CO₂ incubator. Cells were used within passage 10 after primary culture. HTNV strain 76–118 was cultured in Vero E6 cells (ATCC Cat# CRL-1586, RRID:CVCL_0574) in our laboratory and titrated using an immunofluorescence staining assay for HTNV nucleocapsid protein (NP) as previously described (22 (link)). The TCID50 was 10⁵/ml, which was calculated using the Reed-Muench method. The recombinant human IFN-α2a was obtained from PBL Interferon Source (Piscataway, NJ, USA) and dissolved in the buffer provided by the manufacturer (composition not disclosed). HUVEC and A549 cells were infected by incubation with HTNV as indicated moi at 37°C for 60 mins. Subsequently, the virus solution was removed and fresh medium added to the cell culture.

Restriction enzymes were purchased from NEB (New England biolabs, Ipswich, MA, USA). T4 DNA Ligase was purchased from Thermo Scientific (Shanghai, China). Eukaryotic expression vectors pMH3-H, pCApuro-H, pMH3-L, pCApuro-L contained full-length IgG1 H & L chain were preserved in our lab27 (link). Chinese hamster ovary (CHO-s) cell line was purchased from AmProtein (Hangzhou, Zhejiang, China) and cultured in DMEM/F12 medium with 10% (v/v) fetal bovine serum (FBS). HUVEC cells purchased from ScienCell research laboratories (Carlsbad, CA, USA) were c\ultured in ECM medium supplement with 5% (v/v) FBS and 1% (v/v) ECGS (ScienCell, San Diego, CA, USA). Cell culture media and trypsin powder were purchased from Life Technologies (Basel, Switzerland).