Author: van der Schaar, H. M.; Melia, C. E.; van Bruggen, J. A. C.; Strating, J. R. P. M.; van Geenen, M. E. D.; Koster, A. J.; Bárcena, M.; van Kuppeveld, F. J. M.
Title: Illuminating the Sites of Enterovirus Replication in Living Cells by Using a Split-GFP-Tagged Viral Protein Document date: 2016_7_6
ID: 1aptufp6_12
Snippet: In addition to the interaction with GBF1, enterovirus 3A also directly binds to the host factor ACBD3 (acyl coenzyme A [acyl-CoA]-binding protein domain 3), although with the plasmid encoding GFP(S1-10) [i.e., pGFP(S1-10)] and either p3A-myc or p3A(S11aa2)-myc. The next day, cells were fixed and subjected to immunofluorescence analysis. The 3A protein was visualized with an antibody staining for p3A-myc (a primary anti-3A [â£-3A] antibody and a .....
Document: In addition to the interaction with GBF1, enterovirus 3A also directly binds to the host factor ACBD3 (acyl coenzyme A [acyl-CoA]-binding protein domain 3), although with the plasmid encoding GFP(S1-10) [i.e., pGFP(S1-10)] and either p3A-myc or p3A(S11aa2)-myc. The next day, cells were fixed and subjected to immunofluorescence analysis. The 3A protein was visualized with an antibody staining for p3A-myc (a primary anti-3A [â£-3A] antibody and a secondary Alexa Fluor 488-labeled antibody), while p3A(S11aa2) was visualized by the GFP fluorescence as a result of assembly with GFP(S1-10). PI4KB (A) and COP-I (B) were detected with immunofluorescence using a secondary Alexa Fluor 594-labeled antibody. Untransfected cells are indicated with a white asterisk. Nuclei were stained with DAPI. Wide-field images were acquired with an Olympus BX60 fluorescence microscope. Bars, 10 m.
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