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. 2021 Aug 26;12:741839. doi: 10.3389/fimmu.2021.741839

Figure 2.

Figure 2

TRIM65 inhibits NLRP3 inflammasome activation in macrophage. (A) PMA-differentiated and LPS-primed THP-1 cells stably expressing shRNA targeting TRIM65 mRNA were stimulated with MSU, R837 or nigericin. Cleaved IL-1β, activated caspase-1 (P20) in the SN and pro-IL-1β and pro-caspase-1 in the input were analyzed by western blotting. (B-F) LPS-primed BMDMs from wild-type and Trim65-/- mice were stimulated with MSU, ATP, nigericin (B–E), or cytosolic LPS (F). Cleaved IL-1β, activated caspase-1 (P20) in the culture supernatants and pro-IL-1β and pro-caspase-1 in cell lysates were analyzed by western blotting (B). Cleaved IL-1β in the culture supernatants was assayed by ELISA (C–F). Data are representative of three independent experiments of duplicate biological repeats (A, B). SN, media supernatants; Input, cell extracts. Student’s t-test, ***P < 0.001, ****P < 0.0001. Data are shown as the means ± SEM of three independent experiments (C–F).