2 bromohexadecanoic acid 2 bp

Anti-HA antibody (#3F10) was purchased from Roche Diagnostics; anti-CERT (#ab72536) from Abcam; chicken antibody against VAP (Kumagai et al., 2014 (link)) anti-TGOLN2/TGN46 (#A304-434A) from Bethyl Laboratories; anti-TOMM20 (#WH0009804M1) from Sigma-Aldrich; anti-Lamp2 (#sc-18822) from Santa Cruz; anti-LBPA (#z-PLBPA) from Echelon Bioscience; anti-EEA1 (#610457) from BD Transduction Laboratories; anti-catalase (#D4P7B) and anti-Rab11a (#2413) from Cell Signaling Technology; anti-Hrs (#10390-1-AP) from Proteintech; anti-GFP (#04404-84) from Nacalai Tesque, and anti-GAPDH (016-25523) from Fujifilm Wako Pure Chemical Corporation. 2-Bromohexadecanoic acid (2-BP, #M1177) was purchased from Sigma-Aldrich. Lipid dye II (#LD02) was from Dojindo Laboratories. Lysenin was a gift from Dr. Sekizawa (Zenyaku Kogyo). GFP-lysenin was a gift from Dr. Kobayashi (Riken). [¹⁴C(U)]L-serine (156 mCi/mmol, #MC265) was from Moravec.

BHK21 cells were maintained in MEM alpha medium (Cytiva; Hyclone, Marlborough, MA) containing 10% FBS (Cytiva; Hyclone, Marlborough, MA), 1× Penicillin Streptomycin (PS) solution (100 IU/mL Penicillin and 100 μg/mL Streptomycin, Corning, New York), and 1× HEPES buffer (Biological Industries, Israel). HEK293T/17 cells and Vero E6 cells were maintained in 10% FBS in DMEM/High glucose medium (Cytiva; Hyclone, Marlborough, MA) with 1× PS solution. Calu-3 cells were maintained in 10% FBS in MEM medium (ThermoFisher, Waltham, MA) with 1× PS solution. Human ACE2 (hACE2) overexpression in BHK21 cells was introduced by the lentiviral vector carrying the hACE2 gene as described previously (Su et al., 2020 (link)). SARS-CoV-2 virus (hCoV-19/Taiwan/4/2020) was kindly provided by the Taiwan Centers for Disease Control and was amplified in Vero E6 cells in M199 medium with 2 μg/mL TPCK-trypsin. All SARS-CoV-2 experiments were performed in a biosafety level 3 (BSL-3) laboratory. 2-Bromohexadecanoic acid (2BP) was purchased from Sigma-Aldrich (Darmstadt, Germany).

Dulbecco’s Modified Eagle Medium (DMEM), Roswell Park Memorial Institute 1640 Medium (RPMI), penicillin, streptomycin, and L-glutamine were obtained from Gibco^TM (Thermo Fisher Scientific, Waltham, MA, USA). Fetal calf serum (FCS), β-mercaptoethanol (2-ME), polybrene, Accutase® solution, brefeldin A (BFA), monensin (MN), 2-bromohexadecanoic acid (2-BP), lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA), and dexamethasone (Dex) were purchased from Sigma-Aldrich (St. Louis, Missouri).