MARC-145 and HEK 293FT cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Carlsbad, CA, USA) with 10% FBS (Invitrogen, CA, USA). Primary porcine pulmonary alveolar macrophages (PAMs) were obtained from specific-pathogen-free (SPF) pigs as previously described [43 (link)] and cultured in RPMI1640 medium (Invitrogen, CA, USA) containing 10% fetal bovine serum (FBS) (Invitrogen, CA, USA). The Chinese highly pathogenic PRRSV strain JXwn06 infectious cDNA clone was described previously [31 (link)].
Mouse anti-FLAG monoclonal antibody (F1804), mouse anti-HA monoclonal antibody (H3663), mouse anti-β-actin monoclonal antibody (A5441), and rabbit anti-Myc polyclonal antibody (C3956) were all purchased from Sigma-Aldrich (St. Louis, MO, USA). Horseradish peroxidase (HRP)-conjugated goat anti-mouse polyclonal antibodies and HRP-conjugated goat anti-rabbit polyclonal antibodies were purchased from ZSGB-Bio (Beijing, China). All the restriction enzymes used in this study were from New England Biolabs Inc. (Beverly, MA, USA).
Mouse anti-FLAG monoclonal antibody (F1804), mouse anti-HA monoclonal antibody (H3663), mouse anti-β-actin monoclonal antibody (A5441), and rabbit anti-Myc polyclonal antibody (C3956) were all purchased from Sigma-Aldrich (St. Louis, MO, USA). Horseradish peroxidase (HRP)-conjugated goat anti-mouse polyclonal antibodies and HRP-conjugated goat anti-rabbit polyclonal antibodies were purchased from ZSGB-Bio (Beijing, China). All the restriction enzymes used in this study were from New England Biolabs Inc. (Beverly, MA, USA).