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. 2022 Feb 25;13:846384. doi: 10.3389/fimmu.2022.846384

Figure 3.

Figure 3

HSC inhibit the activation of NLRP3 inflammasome. J774A.1, THP-1 cells treated with HSC for 4 h, LPS (1 μg/mL) was added to stimulate for 6 h, followed by ATP (5 mM) co-culture for 30 min; (A, B) NLRP3 inflammasome related proteins NLRP3, ASC, caspase1, caspase-1p20, IL-1β, and IL-1β p17 were determined by western blotting in J774A.1 (A), THP-1 (B) cells; (C, D) The level of IL-1β was determined by ELISA in J774A.1 (C), THP-1(D); (E) ASC activation was determined by immunofluorescence in THP-1 cells; (F) Cleaved-caspase-1 activation was determined by immunofluorescence in J774A.1 cells; (G) Plasmids of EGFP-NLRP3 were transfected into J774A.1 cells for 48 h, NLRP3 activation was determined by immunofluorescence; (H) The proteins were immunoprecipitated with ASC using magnetic beads, and the immunocomplexes were identified by western blotting. (I) Using magnetic beads, the obtained proteins were immunoprecipitated with NLRP3, and immunocomplexes were evaluated by western blotting; (Scale bar = 20 μm); # P < 0.001 compared to control alone group; * P < 0.05, ** P < 0.01, and *** P < 0.001, versus the LPS+ATP group, (n=3).