Fig. 4

SARS-CoV-2 NSP13 interacts with STAT1 to prevent it from being phosphorylated by JAK1. A, B NSP13 interacts with STAT1 but not JAK1. HEK293T cells were transfected with vectors expressing the indicated proteins. Immunoprecipitation (IP) was performed 48 h post transfection. NSP13 was immunoprecipitated with anti-FLAG or anti-V5. Precipitates were probed with anti-V5, anti-FLAG or anti-myc. C NSP13 does not impede complex formation between JAK1 and STAT1. HEK293T cells were transfected with plasmids expressing the indicated proteins. Immunoprecipitation were performed at 48 h post transfection. STAT1 was immunoprecipitated with anti-myc. Precipitates were probed with anti-V5 and anti-myc. D NSP13 blocks STAT1 phosphorylation by JAK1. HEK293T cells was transfected with expression plasmid for V5-tagged NSP13. At 48 h post-transfection, NSP13 protein was immunoprecipitated with anti-V5. The precipitates were incubated with recombinant STAT1 and JAK1 in the presence of 10 mM ATP for 30 min at 30 °C. Phospho-STAT1 (pSTAT1) was probed with anti-pSTAT1 antibodies. Results were representative of three independent experiments