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. Author manuscript; available in PMC: 2020 Dec 2.
Published in final edited form as: Cell Rep. 2020 Nov 17;33(7):108405. doi: 10.1016/j.celrep.2020.108405

Figure 1. RACK1 Interacts with NLRP3 and NEK7.

Figure 1.

(A) Mass spectrometry analysis of NLRP3, RACK1, and NEK7 peptides after purification of NLRP3 complexes.

(B and C) Tagged NLRP3 (NLRP3-SFP) or NEK7 (NEK7-SFP) was immunoprecipitated (IP) with anti-FLAG antibody from iBMDM treated with LPS (200 ng Ml−1, 4 h) alone or with LPS plus ATP (5 mM, 30 min) and was immunoblotted with the indicated antibodies. EV, empty vector.

(D) BMDM was stimulated with LPS (200 ng mL−1, 4 h) alone or with LPS plus ATP (5 mM, 30 min). Cell lysates were immunoprecipitated and immunoblotted with the indicated antibodies.

(E) NLRP3-SFP was co-expressed with HA-tagged RACK1 in HEK293T cells, pulled down, and analyzed by immunoblotting. HA, hemagglutinin.

(F) FLAG-tagged, full-length or truncated NLRP3 was co-expressed with HA-tagged RACK1 in HEK293T cells, immunoprecipitated, and analyzed by immunoblotting. (Δpyrin, pyrin domain deleted; ΔLRR, leucine-rich repeats deleted; pyrin, pyrin domain only; NOD, NOD domain only; LRR, leucine-rich repeat only). Results are representative of three independent experiments.