Journal: iScience

Article Title: Grass carp reovirus VP56 and VP35 induce formation of viral inclusion bodies for replication

doi: 10.1016/j.isci.2023.108684

Figure Lengend Snippet: GCRV-II NS38 interacts with VP1, VP56, and VP35 (A) 293T cells were transfected with pEGFP-NS38, together with pFlag-VP1, pFlag-VP2, pFlag-VP3, pFlag-NS79, pFlag-VP5, pFlag-VP4, pFlag-VP56, pFlag-VP41, pFlag-VP6, pFlag-VP35, or p3XFlag-CMV-14 (control). The cells were collected at 24 h post transfection, followed by Co-IP assay with anti-Flag antibody. Then the immunoprecipitates and cell lysates were analyzed with anti-Flag and anti-EGFP antibodies. (B) 293T cells were transfected with pEGFP-NS38, together with pDsRed-VP1, pDsRed-VP56, or pDsRed-VP35. At 24 h post transfection, the cells were fixed and stained with DAPI. The samples were detected using the confocal microscope. Scale bar: 10 μm.

Article Snippet: Anti-Flag Magnetic Beads , MedChemExpress , Cat#HY-K0207.

Techniques: Transfection, Control, Co-Immunoprecipitation Assay, Staining, Microscopy

Journal: iScience

Article Title: Grass carp reovirus VP56 and VP35 induce formation of viral inclusion bodies for replication

doi: 10.1016/j.isci.2023.108684

Figure Lengend Snippet: GCRV VP56 interacts with VP1, NS79, VP5, NS38, and VP35 (A) 293T cells were transfected with pEGFP-VP56, together with pFlag-VP1, pFlag-VP2, pFlag-VP3, pFlag-NS79, pFlag-VP5, pFlag-VP4, pFlag-VP41, pFlag-VP6, pFlag-NS38, pFlag-VP35, or p3XFlag-CMV-14(control). The cells were collected at 24 h post transfection, followed by Co-IP assay with anti-Flag antibody. Then the immunoprecipitates and cell lysates were analyzed with anti-Flag and anti-EGFP antibodies. (B) 293T cells were transfected with pEGFP-VP56, together with pDsRed-VP1, pDsRed-NS79, pDsRed-VP5, pDsRed-NS38, or pDsRed-VP35. At 24 h post transfection, the cells were fixed and stained with DAPI. The samples were detected using confocal microscope. Scale bar: 10 μm.

Article Snippet: Anti-Flag Magnetic Beads , MedChemExpress , Cat#HY-K0207.

Techniques: Transfection, Control, Co-Immunoprecipitation Assay, Staining, Microscopy

Journal: iScience

Article Title: Grass carp reovirus VP56 and VP35 induce formation of viral inclusion bodies for replication

doi: 10.1016/j.isci.2023.108684

Figure Lengend Snippet: GCRV VP35 interacts with VP1, VP2, VP3, VP4, VP56, and NS38 (A) 293T cells were transfected with pEGFP-VP35, together with pFlag-VP1, pFlag-VP2, pFlag-VP3, pFlag-NS79, pFlag-VP5, pFlag-VP4, pFlag-VP56, pFlag-VP41, pFlag-VP6, pFlag-NS38, or p3XFlag-CMV-14(control). The cells were collected at 24 h post transfection, followed by Co-IP assay with anti-Flag antibody. Then the immunoprecipitates and cell lysates were analyzed with anti-Flag and anti-EGFP antibodies. (B) 293T cells were transfected with pEGFP-VP35, together with pDsRed-VP1, pDsRed-VP2, pDsRed-VP3, pDsRed-VP4, pDsRed-VP56, or pDsRed-NS38. At 24 h post transfection, the cells were fixed and stained with DAPI. The samples were detected using confocal microscope. Scale bar: 10 μm.

Article Snippet: Anti-Flag Magnetic Beads , MedChemExpress , Cat#HY-K0207.

Techniques: Transfection, Control, Co-Immunoprecipitation Assay, Staining, Microscopy

Journal: iScience

Article Title: Grass carp reovirus VP56 and VP35 induce formation of viral inclusion bodies for replication

doi: 10.1016/j.isci.2023.108684

Figure Lengend Snippet: N-terminal domain of VP56 is responsible for the VIBs formation and interaction with the recruited viral proteins (A) The schematic shows the plasmids expressing different VP56 truncated mutants. (B) GCO cells were transfected with pEGFP-VP56-N, pEGFP-VP56-M, or pEGFP-VP56-C. At 24 h post transfection, the cells were fixed and stained with DAPI. The samples were detected using confocal microscope. Scale bar: 10 μm. (C) 293T cells were transfected with pEGFP-VP56-N, together with pMyc-VP56-N, pFlag-VP1, pFlag-NS79, pFlag-VP5, pFlag-NS38, or pFlag-VP35. The cells were collected at 24 h post transfection, followed by Co-IP assay with anti-Myc or anti-Flag antibody. Then the immunoprecipitates and cell lysates were analyzed with anti-EGFP, anti-Myc, and anti-Flag antibodies.

Article Snippet: Anti-Flag Magnetic Beads , MedChemExpress , Cat#HY-K0207.

Techniques: Expressing, Transfection, Staining, Microscopy, Co-Immunoprecipitation Assay

Journal: iScience

Article Title: Grass carp reovirus VP56 and VP35 induce formation of viral inclusion bodies for replication

doi: 10.1016/j.isci.2023.108684

Figure Lengend Snippet: Middle domain of VP35 is responsible for the VIBs formation and interaction with the recruited viral proteins (A) The schematic shows the plasmids expressing different VP35 truncated mutants. (B) GCO cells were transfected with pEGFP-VP35-N, pEGFP-VP35-M, or pEGFP-VP35-C. At 24 h post transfection, the cells were fixed and stained with DAPI. The samples were detected using confocal microscope. Scale bar: 10 μm. (C) 293T cells were transfected with pEGFP-VP35-M, together with pMyc-VP35-M, pFlag-VP1, pFlag-VP2, pFlag-VP3, pFlag-VP4, pFlag-VP56, or pFlag-NS38. The cells were collected at 24 h post transfection, followed by Co-IP assay with anti-Myc or anti-Flag antibody. Then the immunoprecipitates and cell lysates were analyzed with anti-EGFP, anti-Myc, and anti-Flag antibodies.

Article Snippet: Anti-Flag Magnetic Beads , MedChemExpress , Cat#HY-K0207.

Techniques: Expressing, Transfection, Staining, Microscopy, Co-Immunoprecipitation Assay

Journal: iScience

Article Title: Grass carp reovirus VP56 and VP35 induce formation of viral inclusion bodies for replication

doi: 10.1016/j.isci.2023.108684

Figure Lengend Snippet:

Article Snippet: Anti-Flag Magnetic Beads , MedChemExpress , Cat#HY-K0207.

Techniques: Virus, Recombinant, Transfection, Lysis, Magnetic Beads, Plasmid Preparation, Software

Journal: eLife

Article Title: In silico screening by AlphaFold2 program revealed the potential binding partners of nuage-localizing proteins and piRNA-related proteins

doi: 10.7554/eLife.101967

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , anti-FLAG magnetic beads , MBL , Cat# M185-11R , .

Techniques: Recombinant, Plasmid Preparation, Magnetic Beads, Transfection, Western Blot, Software

Journal: Journal of Virology

Article Title: Biochemical analysis of the host factor activity of ZCCHC14 in hepatitis A virus replication

doi: 10.1128/jvi.00057-24

Figure Lengend Snippet: (A) Immunoprecipitation of 293T lysate expressing HA-ZCCHC14 with purified recombinant FLAG-TENT4A or FLAG-TENT4B proteins. BSA is used as a negative control. (B) IP as in (A) with indicated ZCCHC14 constructs. (C) IP as in (B) with RNase 1 treatment. (D) (left) Diagram depicting deletions made in the Z14-N fragment and (right) their interaction with TENT4 in the co-IP assay. (E) (left) Diagram depicting deletions made in the Z14-C fragment and (right) their interaction with TENT4 in the co-IP assay.

Article Snippet: For this purpose, 293T lysates expressing HA-tagged ZCCHC14 fragments were incubated with purified recombinant FLAG-tagged TENT4 protein fragments (aa 226–558 of TENT4A or aa 186–518 of TENT4B) representing the catalytic domains of these noncanonical polymerases (kindly provided by Arbutus Biopharma) or a bovine serum albumin (BSA) control and immunoprecipitated with anti-FLAG magnetic beads.

Techniques: Immunoprecipitation, Expressing, Purification, Recombinant, Negative Control, Construct, Co-Immunoprecipitation Assay