Vero cells grown on microscope coverslips placed in 6‐well tissue culture plates were mock infected or infected with PEDV at a multiplicity of infection (MOI) of 0.1. The virus‐infected cells were cultured until 24 hr, fixed with 4% paraformaldehyde for 10 min at room temperature (RT) and permeabilized with 0.2% Triton X‐100 in PBS at RT for 10 min. The cells were blocked with 1% bovine serum albumin (BSA) in PBS for 30 min at RT and then incubated for 2 hr with a monoclonal antibody (MAb) specific for PEDV N protein (ChoogAng Vaccine Laboratories). After being washed five times with PBS, the cells were incubated for 1 hr at RT with a goat anti‐mouse secondary antibody conjugated to Alexa Fluor 488 (Invitrogen) followed by counterstaining with 4′,6‐diamidino‐2‐phenylindole (DAPI; Sigma). The coverslips were mounted onto glass microscope slides using mounting buffer and the stained cells were visualized using a fluorescence Leica DM IL LED microscope (Leica).
Copyright and License information: ©2019 Blackwell Verlag GmbH This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.
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