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. 2021 Apr 13;12(2):e00065-21. doi: 10.1128/mBio.00065-21

FIG 6.

FIG 6

SARS-CoV-2 ORF6 represses reporter expression and copurifies with relatively more Rae1-Nup98 than SARS-CoV ORF6. (A) Comparison between the amino acid sequences of ORF6 of SARS-CoV and ORF6 of SARS-CoV-2. Residues differing between the two viruses are highlighted in red. The residue (Met58) implicated in binding the Rae1·Nup98 complex is highlighted in blue. (B) 293T cells were transiently transfected with GFP-tagged constructs and mCherry and visualized 24 h after transfection. Bars, 100 μm. (C) Cells transfected with SARS-CoV-2 ORF6 showed significantly reduced mCherry expression compared to those transfected with SARS-CoV ORF6. (D) Western blotting showed decreased expression of SARS-CoV-2 ORF6 compared to SARS-CoV ORF6 in 293T cells. (E) Affinity purification of GFP-tagged constructs demonstrates both ORF6 of SARS-CoV and ORF6 of SARS-CoV-2 interact with Rae1 and Nup98. (F and G) Densitometry shows SARS-CoV-2 ORF6 copurifies with relatively more Rae1 (F) and Nup98 (G) compared to SARS-CoV ORF6. *, P < 0.05; **, P < 0.01.