Journal: Journal of Virology

Article Title: Porcine Sapelovirus Uses α2,3-Linked Sialic Acid on GD1a Ganglioside as a Receptor

doi: 10.1128/JVI.02449-15

Figure Lengend Snippet: PSV binding and infection require carbohydrate moieties. (A) The binding of AF-594-labeled PSV (MOI, 1,000 PFU/cell) was examined by confocal microscopy after pretreating LLC-PK cells with NaIO 4 . (B) Binding of [ 35 S]methionine-cysteine-labeled PSV, EV70, FCV, or CVB3 (50,000 cpm) was measured by liquid scintillation counting after treating cell or not with NaIO 4 . The levels of bound virus were expressed as a percentage of the value for the mock-treated, virus-infected control. (C) The effect of NaIO 4 pretreatment of permissive cells on the infectivity of PSV (MOI, 1 PFU/cell) was assessed by immunofluorescence using monoclonal antibody specific for PSV capsid protein at 15 h postinfection. (D) The levels of PSV, EV70, FCV, or CVB3 antigen-positive cells expressed as percentages of mock-treated, virus-infected cells were quantified from three independent fields of view. All experiments were performed three independent times. Panels A and C show representative sets of results. The scale bars corresponded to 20 μm. Error bars indicated standard deviations (SD) from triplicate experiments. *, P < 0.05; **, P < 0.005.

Article Snippet: Enterovirus 70 (EV70) strain J670/71 (ATCC) was propagated in WI-38 cells ( ).

Techniques: Binding Assay, Infection, Labeling, Confocal Microscopy, Virus, Control, Immunofluorescence

Journal: Journal of Virology

Article Title: Porcine Sapelovirus Uses α2,3-Linked Sialic Acid on GD1a Ganglioside as a Receptor

doi: 10.1128/JVI.02449-15

Figure Lengend Snippet: PSV binding and infection are blocked by N -acetylneuraminic acid (NANA). (A) The effect of a soluble form of NANA on the binding of AF-594-labeled PSV (MOI, 1,000 PFU/cell) to LLC-PK cells was examined by confocal microscopy. (B) The effect of soluble NANA, Gal, or 6′-SL on the binding of [ 35 S]methionine-cysteine-labeled PSV, EV70, FCV, or CVB3 (50,000 cpm) to each corresponding cell was measured by liquid scintillation and expressed as a percentage of the value for the mock-treated, virus-infected control. (C) The effect of NANA of permissive cells on the infectivity of PSV (MOI, 1 PFU/cell) was assessed by immunofluorescence using monoclonal antibody specific for PSV capsid protein at 15 h postinfection. (D) The levels of PSV, EV70, FCV, or CVB3 antigen-positive cells expressed as percentages of the mock-treated, virus-infected control were quantified from three independent fields of view. All experiments were performed three independent times. Panels A and C show representative sets of results. Scale bars corresponded to 20 μm. Error bars indicate SD from triplicate experiments. *, P < 0.05; **, P < 0.005.

Article Snippet: Enterovirus 70 (EV70) strain J670/71 (ATCC) was propagated in WI-38 cells ( ).

Techniques: Binding Assay, Infection, Labeling, Confocal Microscopy, Virus, Control, Immunofluorescence

Journal: Journal of Virology

Article Title: Porcine Sapelovirus Uses α2,3-Linked Sialic Acid on GD1a Ganglioside as a Receptor

doi: 10.1128/JVI.02449-15

Figure Lengend Snippet: PSV binding and infection require terminal SA. (A) Binding of AF-594-labeled PSV (MOI, 1,000 PFU/cell) was examined by confocal microscopy after pretreating LLC-PK cells with NA or SS. (B) The effect of pretreament of NA or SS to each corresponding cell on the binding of [ 35 S]methionine-cysteine-labeled PSV, EV70, FCV, or CVB3 (50,000 cpm) was examined by liquid scintillation counting and expressed as a percentage of the value for the mock-treated, virus-infected control. (C) The effect of NA or SS pretreatment of permissive cells on the infectivity of PSV (MOI, 1 PFU/cell) was assessed by immunofluorescence using monoclonal antibody specific for PSV capsid protein at 15 h postinfection. (D) The levels of PSV, EV70, FCV, or CVB3 antigen-positive cells expressed as percentages of mock-treated, virus-infected cells were quantified from three independent fields of view. All experiments were performed three independent times. Panels A and C show representative sets of results. The scale bars correspond to 20 μm. Error bars indicate SD from triplicate experiments. *, P < 0.05; **, P < 0.005.

Article Snippet: Enterovirus 70 (EV70) strain J670/71 (ATCC) was propagated in WI-38 cells ( ).

Techniques: Binding Assay, Infection, Labeling, Confocal Microscopy, Virus, Control, Immunofluorescence

Journal: Journal of Virology

Article Title: Porcine Sapelovirus Uses α2,3-Linked Sialic Acid on GD1a Ganglioside as a Receptor

doi: 10.1128/JVI.02449-15

Figure Lengend Snippet: PSV binding and infection require terminal α2,3-linked SA. (A) Binding of AF-594-labeled PSV (MOI, 1,000 PFU/cell) was examined by confocal microscopy after pretreating LLC-PK cells with MAL or SNL. (B) The effect of pretreament of MAL or SNL to each corresponding cell on the binding of [ 35 S]methionine-cysteine-labeled PSV, EV70, FCV, and CVB3 strains (50,000 cpm) was examined by liquid scintillation counting and expressed as a percentage of the value for the mock-treated, virus-infected control. (C) The effect of MAL or SNL pretreatment of permissive cells on the infectivity of PSV (MOI, 1 PFU/cell) was assessed by immunofluorescence using monoclonal antibody specific for PSV capsid protein at 15 h postinfection. (D) The levels of PSV, EV70, FCV, or CVB3 antigen-positive cells expressed as percentages of the mock-treated, virus-infected cells were quantified from three independent fields of view. All experiments were performed three independent times. Panels A and C show representative sets of results. The scale bars correspond to 20 μm. Error bars indicate SD from triplicate experiments. *, P < 0.05; **, P < 0.005.

Article Snippet: Enterovirus 70 (EV70) strain J670/71 (ATCC) was propagated in WI-38 cells ( ).

Techniques: Binding Assay, Infection, Labeling, Confocal Microscopy, Virus, Control, Immunofluorescence