Alexa fluor 594 goat anti rabbit immunoglobulin g h l

To analyze the colocalization of ISG15 with PPRV proteins, EECs after cotransfection with plasmids or infection with PPRV were fixed with 4% paraformaldehyde for 30 min at room temperature and washed three times with PBS for 6 min each. The cells were permeabilized by prechilled methanol at −20°C and incubated at the same temperature for 10 min. The cells were blocked with 5% bovine serum albumin and 0.3% Triton X-100 at room temperature for 2 h. Primary antibodies and secondary antibodies (Alexa Fluor 594 goat anti-rabbit immunoglobulin G [H+L] and Alexa Fluor 488 goat anti-mouse immunoglobulin G [H+L]; Invitrogen) were then added. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen). Images were taken with a Zeiss LSM880 confocal microscope (Zeiss).

To analyze the colocalization of ISG15 with PPRV proteins, EECs after cotransfection with plasmids or infection with PPRV were fixed with 4% paraformaldehyde for 30 min at room temperature and washed three times with PBS for 6 min each. The cells were permeabilized by prechilled methanol at −20°C and incubated at the same temperature for 10 min. The cells were blocked with 5% bovine serum albumin and 0.3% Triton X-100 at room temperature for 2 h. Primary antibodies and secondary antibodies (Alexa Fluor 594 goat anti-rabbit immunoglobulin G [H+L] and Alexa Fluor 488 goat anti-mouse immunoglobulin G [H+L]; Invitrogen) were then added. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen). Images were taken with a Zeiss LSM880 confocal microscope (Zeiss).