Wb and ip lysis buffer

Phenyl methane sulfonyl fluoride (PMSF; #ST505), proteinase K (#ST533), tritonX-100 (#P0096), WB and IP lysis buffer (#P0013), RIPA lysis buffer (#P0013D), DAPI staining solution (#C1005), 4% paraformaldehyde (PFA; #P0099), BeyoECL plus (#P0018S), enhanced BCA protein assay kit (#P0010S), and enhanced cell counting kit-8 (CCK-8; #C0041) were produced from Beyotime Biotechnology (Shanghai, China). TurboFect transfection reagent (#R0533), GST protein interaction pull-down kit (#21516), and OPP protein synthesis assay kits (#C10457) were obtained from Thermo Fisher Scientific. RNAiso plus reagent (#9108Q), viral RNA/DNA extraction kit (#9766), Prime Script RT reagent kit (#RR047B), and TB Green fast qPCR mix (#RR430A) were obtained from TAKARA (Dalian, China). Dual-luciferase reporter assay system (#E1910) was obtained from Promega (Madison, USA). Puromycin (#P8833), cholesterol (#C4951), cycloheximide (#239763-M), hexadimethrine bromide (#H9268), and L-α-Phosphatidylcholine (#P3556) were obtained from Sigma Aldrich. Chemically defined lipid concentrate (#11905-031) and advanced DMEM (#12491-015) were obtained from Invitrogen (Carlsbad, USA).

The total proteins were extracted from cells with WB and IP lysis buffer (Beyotime Biotechnology, Shanghai, China). Equal amounts of proteins were loaded in SDS-polyacrylamide gels, and transferred to PVDF membranes (Millipore, USA). After blocking with 5% BSA at room temperature for 2 h, the membranes were incubated with primary antibodies at 4°C overnight and then incubated with secondary antibodies for 1.5 h at room temperature. Proteins were then measured by an enhanced chemiluminescence system (ECL) reagent (KeyGEN BioTECH, China). Band intensity was quantified by densitometry analysis using Image-Pro Plus 4.5 software (Rockville, MD, United States).