Fig. 6

2N protein inhibits RIG-I ubiquitination through TRIM25. a HEK293T cells in six-well plate were transfected with HA-Ub and Flag-RIG-I plasmids as indicated, together with an increasing GST-SARS-CoV-2 N (GST-2N) plasmid. At 24 hpt, anti-Flag immunoprecipitates were analyzed by immunoblot with anti-Ub, HA, Flag, and GST antibodies. b HEK293T cells in 24-well plate were transfected with Flag-RIG-I, or Flag-RIG-I and HA-Ub, or Flag-RIG-I, HA-Ub, and 0.25 μg GST-2N plasmids. At 24 hpt, the cells were subjected to immunofluorescence with anti-Flag and HA antibodies. c HEK293T cells in six-well plate were transfected with the plasmids as indicated and 1 μg 2N plasmid, anti-Flag immunoprecipitates were analyzed by immunoblot. d sg25 and WT cells in six-well plate were transfected with the indicated plasmids and 1 μg 2N plasmid, anti-Flag immunoprecipitates were analyzed by immunoblot