Fig. 2

Confirmation of protein interactions by co-immunoprecipitation. The coding sequences of each viral proteins fused with Flag or HA tag were cloned into LPC vector, respectively. The combination of two proteins with indicated tags was expressed in HEK293T cells, and the cell lysates were collected for co-immunoprecipitation with Flag agarose. The samples from co-immunoprecipitation were examined with WB, and the two proteins were detected with Flag and HA antibodies. ORF8 was fused with EGFP for an increased expression level. Four self-interactions, ORF8 (A), N (B), Nsp15 (C), and Nsp5 (D), and Four interactions between various viral proteins, Nsp14-Nsp10 (E), Nsp16-Nsp10 (F), Nsp3.1-N (G), and Nsp8-Nsp12 (H) were confirmed using co-immunoprecipitation. Data represent one of 3 independent experiments with similar results