Skip to main content
. 2020 Nov 5;102(1):jgv001513. doi: 10.1099/jgv.0.001513

Fig. 1.

Fig. 1.

SARS-CoV-2 nsp1 inhibits host-protein synthesis. (a) Luciferase activities in HEK-293T cells transfected with 0.5 µg of either PCAGGS-SARS-CoV-2 nsp1 (SARS-CoV-2 nsp1), PCAGGS-SARS-CoV nsp1 (SARS-CoV nsp1) or PCAGGS (Mock) together with 0.2 µg of the indicated reporter plasmid (pRL-TK or pRL-SV40) were determined at 36 h post-transfection after standardization with cells expressing SARS-CoV-2 nsp1. Error bars show the standard deviation (SD) of the results from three independent experiments. Cell extracts were also subjected to Western-blot analysis using an anti-HA antibody (top) or anti-β-actin antibody (bottom). ***, P <0.001. (b) HEK-293T cells were cotransfected with pRL-SV40 encoding the Rluc reporter gene downstream of the SV40 promoter and one of the following plasmids: PCAGGS (mock), PCAGGS-SARS-CoV-2 nsp1 (SARS-CoV-2 nsp1) or PCAGGS-SARS-CoV nsp1 (SARS-CoV nsp1), respectively. At 24 h post-transfection, the cells were lysed and subjected to real-time quantitative PCR analysis. The values of SARS-CoV-2 nsp1 and SARS-CoV nsp1 were normalized to those of the untreated empty vector (PCAGGS) control, which was set to 1 (n=3). Asterisks indicate statistical significance calculated based on Student’s t-test. ns, not significant; **, P <0.01. (c) HEK-293T cells were transfected with 2.5 µg of either PCAGGS-SARS-CoV-2 nsp1 (SARS-CoV-2 nsp1), PCAGGS-SARS-CoV nsp1 (SARS-CoV nsp1) or PCAGGS (Mock) plasmid. At 36 h post-transfection, the cells were pulsed with 3 µm puromycin for 1 h at 37 °C and then subjected to Western-blot analysis using an anti-puromycin antibody (top), anti-HA antibody (middle), or anti-β-actin antibody (bottom). (d and e) The grey scale values of the proteins were analysed by ImageJ (n=3). *, P <0.05; **, P <0.01.