Fig 1.
SARS-CoV-2 Mpro cleaves MAGED2 at Gln-263. (A) Putative Mpro cleavage sites in SARS-CoV-2 non-structural proteins and MAGED2. (B and C) HEK293T cells were co-transfected with human MAGED2 or Q263N mutant with Flag tag at C-terminal and HA-tagged SARS-CoV-2 Mpro or a proteolytically inactive mutant Mpro (C145A). Lysates from transfected cells were prepared for immunoblotting with antibodies, as indicated. (D) MAGED2 mutant with Flag tag at C-terminal and HA-tagged Mpro were co-expressed in HEK293T cells. Lysates from transfected cells were prepared for immunoblotting with antibodies, as indicated. (E) MAGED2 cleavage assay in vitro. Purified MAGED2 and Mpro wild-type (WT) or C145A mutant proteins were incubated in vitro and analyzed by Coomassie blue staining. One star is MAGED2N, and two stars indicate MAGED2C. (F) A549-hACE2 cells were infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.1 or 1, and immunoblot was performed at 24-h post-infection. Red star indicates cleaved MAGED2. Each experiment was independently repeated three times with similar results, and the representative images are shown.