A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research
Fig 3
Usage of our toolkit antibodies to demonstrate or confirm SARS-CoV-2 protein interactions.
(A) A co-IP was performed using lysates from Vero E6 cells infected with England-02 at MOI 0.1 for 3 days. Using the specific anti-ORF3a antibody described herein, SARS-CoV-2 ORF3a was immunoprecipitated (alongside a preimmune IgG control), and the immune complexes were western blotted for the presence of SARS-CoV-2 spike (S) and ORF3a. (B) As in (A), SARS-CoV-2 nsp13 (or IgG control) was immunoprecipitated and the immune complexes were probed for nsp13 and nsp11/12 by WB. (C) As in (A, B), SARS-CoV-2 N (or IgG control) was immunoprecipitated and the immune complexes were probed for matrix (M) and N by WB. co-IP, co-immunoprecipitation; IB, immunoblotting; IgG, immunoglobulin G; MOI, multiplicity of infection; SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus 2; WB, western blotting.
doi: https://doi.org/10.1371/journal.pbio.3001091.g003