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. Author manuscript; available in PMC: 2023 Apr 15.
Published in final edited form as: J Immunol. 2022 Mar 30;208(8):2029–2036. doi: 10.4049/jimmunol.2101154

FIGURE 5.

FIGURE 5.

The basic property of R121NEK7 is critical for NLRP3 binding and inflammasome activation. (A) Flag-tagged NLRP3 was co-expressed with HA-tagged wild-type or mutant NEK7 in HEK 293T cells. Cell lysates were immunoprecipitated with anti-Flag antibody and immunoblotted with indicated antibodies. (B) NEK7 KO iBMDMs were reconstituted with HA-tagged mouse wild-type or mutant NEK7 by lentiviral transduction. Macrophages were stimulated with 5 μM nigericin for 1 h after LPS priming. Mixtures of cell lysates and supernatants were immunoblotted with indicated antibodies. (C) Measurement of IL-β in the supernatants from nigericin-stimulated macrophages by ELISA. EV, empty vector. Actin was used as an internal control. Data are representative of three experiments. Error bars in C show the SEM of triplicate wells. *p < 0.05.