Fig. 1
From: SARS-CoV-2 hijacks cellular kinase CDK2 to promote viral RNA synthesis
CDKs involve in the activity of SARS-CoV-2 RdRp. a Flag-tagged nsp7, nsp8, nsp12, or Flag-vector were co-transfected in HEK293T cells. Anti-Flag M2 affinity gel was used for Co-Immunoprecipitation and Western blot analysis was performed with the indicated antibodies. b The immunoprecipitated samples were separated by SDS-PAGE followed by coomassie blue staining. Asterisk (*) indicates the band for mass spectrometry analysis. c The top 20 candidate genes indicated by mass spectrometry. d, e HEK293T cells expressing CoV-Gluc, nsp12, nsp7, nsp8 plasmid DNA at the ratio of 1:10:30:30 (d) or control vector and CoV-Gluc (e) were transfected with CDK1 or CDK2 siRNA (three siRNAs per gene) for 48 h. Then the Gluc activity was measured in the supernatants. The CDK1/2 knockdown were determined by western blot analysis. f, g HEK293T cells expressing CoV-Gluc, nsp12, nsp7, nsp8 plasmid DNA at the ratio described above (f) or control vector and CoV-Gluc (g) were transfected with CDK2/CyclinA plasmids or CDK1/CyclinB plasmids for 48 h. Gluc activity was measured in the supernatants. CDK1/2 overexpression was detected by qRT-PCR. The experiments was performed at least three times in d–g, and data are presented as mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001 and ns not significant (two-tailed unpaired Student’s t-test)
