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. 2011 Dec;85(24):13019–13026. doi: 10.1128/JVI.05942-11

Fig. 4.

Fig. 4.

Interaction of the MV V protein with NLRP3. (A) HEK293T cells were transfected with expression plasmids encoding HA-tagged V protein and either Flag-tagged NLPR3, ASC, MDA5, or empty vector. Proteins were immunoprecipitated 48 h later with anti-Flag antibody, followed by immunoblot analysis of total lysates (INPUT) and immunoprecipitates (IP) with indicated antibodies (anti-Flag or anti-HA). (B) V, Vn (top and middle), and Vc (bottom) were examined for their interaction with NLRP3 as described for panel A. (C) Vc and Vc with the C272R substitution (VcC272R) were examined for their interaction with NLRP3 as described for panel A. Vc and VcC272R were expressed as fusion proteins with kusabira orange, so that the control HA-empty plasmid directed the production of the kusabira orange peptide, which was detected with anti-HA-tag antibody. (D) PMA-stimulated THP-1/MV-V cells were treated with LPS (5 ng/ml) and ATP (5 mM) or left untreated for 6 h and examined for the localization of NLRP3 and MV V proteins. The data shown are representative of at least three experiments.