Journal: Signal transduction and targeted therapy

Article Title: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) membrane (M) protein inhibits type I and III interferon production by targeting RIG-I/MDA-5 signaling.

doi: 10.1038/s41392-020-00438-7

Figure Lengend Snippet: Fig. 5 The SARS-CoV-2 M protein prevents the formation of the RIG-I–MAVS–TRAF3–TBK1 multiprotein complex. The SARS-CoV-2 M protein inhibits the RIG-I–MAVS (a), MAVS–TBK1 (c), and TRAF3–TBK1 (d) but not MDA-5–MAVS (b) interactions. The HEK293T cells were transfected with the indicated plasmids for 24 h before coimmunoprecipitation with the antibodies against MAVS (a and b) or TBK1 (c and d). The pcDNA6B empty vector was used to balance the total amount of plasmid DNA in the transfection. The input and immunoprecipitates were immunoblotted with the indicated antibodies. Immunoblotting results are representative of two independent experiments. SARS-CoV-2 M protein, SCV2-M

Article Snippet: Antibodies and reagents Rabbit anti-DYKDDDDK tag (D6W5B), rabbit anti-RIG-I (D14G6), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2), and rabbit anti-TRAF3 antibodies (Abs) were obtained from Cell Signaling Technology (USA); the mouse antiMAVS (E-3) and mouse anti-TRAF3 (G-6) antibodies were obtained from Santa Cruz Biotechnology (USA); the mouse anti-actin and rabbit anti-calnexin antibodies were obtained from Proteintech (Wuhan, China); the mouse anti-Flag M2 antibody was obtained from SigmaAldrich (USA); the mouse anti-Myc (9E10) antibody was obtained from Origene (USA); the rabbit anti-GM130 antibody was obtained from Abcam (United Kingdom); the mouse anti-GAPDH antibody (AF0006) was obtained from Beyotime (China); and the mouse anti-HA antibody was obtained from MDL Biotech (China).

Techniques: Transfection, Plasmid Preparation, Western Blot