Journal: Virologica Sinica

Article Title: Autographa Californica Multiple Nucleopolyhedrovirus P48 ( Ac103 ) Is Required for the Efficient Formation of Virus-Induced Intranuclear Microvesicles

doi: 10.1007/s12250-019-00147-8

Figure Lengend Snippet: Western blot analysis of P48 in purified and fractionated virions. BVs and ODVs were purified from vP48:FLAG-infected larvae and fractionated into envelope and nucleocapsid fractions. Immunoblotting was performed with an anti-FLAG antibody to detect FLAG-tagged P48, an anti-VP39 antibody to detect the major capsid protein VP39, an anti-ODV-E25 antibody to detect the BV/ODV envelope-associated protein ODV-E25, and an anti-GP64 antibody to detect the BV envelope-specific protein GP64. NC, nucleocapsid fraction; E, envelope fraction.

Article Snippet: The mouse monoclonal anti-FLAG antibody, mouse monoclonal anti-Myc antibody, and 50% slurry of protein A/G agarose beads conjugated to an anti-Myc antibody or anti-FLAG antibody were purchased from Abmart (Shanghai, China).

Techniques: Western Blot, Purification, Infection

Journal: Virologica Sinica

Article Title: Autographa Californica Multiple Nucleopolyhedrovirus P48 ( Ac103 ) Is Required for the Efficient Formation of Virus-Induced Intranuclear Microvesicles

doi: 10.1007/s12250-019-00147-8

Figure Lengend Snippet: Immunogold localization of P48 in vP48:FLAG-infected cells. Sf9 cells infected with vP48:FLAG at an MOI of 10 TCID50/cell were harvested at 60 h p.i. and processed for immunogold labeling with a mouse monoclonal anti-FLAG antibody. The arrows indicate the immunogold labeling of P48 on the nucleocapsids in the VS (A), nucleocapsid (black arrow) and envelope (white arrow) of ODVs (B), intranuclear microvesicles (C), nuclear envelope (D), and the plasma membrane (E). Cy, cytoplasm; Nu, nucleus; PM, plasma membrane; NE, nuclear envelope. Scale bars, 200 nm.

Article Snippet: The mouse monoclonal anti-FLAG antibody, mouse monoclonal anti-Myc antibody, and 50% slurry of protein A/G agarose beads conjugated to an anti-Myc antibody or anti-FLAG antibody were purchased from Abmart (Shanghai, China).

Techniques: Infection, Labeling, Clinical Proteomics, Membrane

Journal: Virologica Sinica

Article Title: Autographa Californica Multiple Nucleopolyhedrovirus P48 ( Ac103 ) Is Required for the Efficient Formation of Virus-Induced Intranuclear Microvesicles

doi: 10.1007/s12250-019-00147-8

Figure Lengend Snippet: Coimmunoprecipitation analysis of the potential associations between P48 and proteins required for intranuclear microvesicle formation. Sf9 cells were cotransfected twice with transient-expression plasmids (A) or coinfected with recombinant viruses (B) separately expressing P48:FLAG and Myc-tagged Ac75, Ac76, Ac93, or P48. At 36 h p.t. or 36 h p.i., the cells were harvested, and the proteins were immunoprecipitated with protein A/G agarose beads conjugated to an anti-Myc antibody. Western blot analyses were performed with an anti-FLAG antibody to detect the expression (Input, top panels) and coimmunoprecipitation (IP: anti-Myc, top panels) of FLAG-tagged P48 and with an anti-Myc antibody to detect the expression (Input, bottom panels) and immunoprecipitation (IP: anti-Myc, bottom panels) of Myc-tagged viral proteins. Cells cotransfected with pIB-P48:FLAG and pIB-V5/His or cells coinfected with vAcWT-P48:FLAG and vAcWT were used as negative controls. Input, cell lysates; IP: anti-Myc, immunoprecipitation with protein A/G agarose beads conjugated to an anti-Myc antibody. IgH, immunoglobulin heavy chain. IgL, immunoglobulin light chain. The molecular masses (in kDa) of protein standards are noted on the left.

Article Snippet: The mouse monoclonal anti-FLAG antibody, mouse monoclonal anti-Myc antibody, and 50% slurry of protein A/G agarose beads conjugated to an anti-Myc antibody or anti-FLAG antibody were purchased from Abmart (Shanghai, China).

Techniques: Expressing, Recombinant, Immunoprecipitation, Western Blot