Fig. 4

Aldose reductase is a key target in NLRP3 inflammasome activation during epalrestat treatment. A Cell lysates of LPS-primed BMDMs stimulated with or without nigericin. Cells were incubated with epalrestat-sepharose for 12 h and the proteins were pulled down with Sepharose beads. B HEK-293 T cells were transfected with Flag-Vector or Flag-NLRP3 for 24 h and then treated with or without epalrestat. Immunoprecipitation was performed using anti-Flag M2 agarose beads. The results of the immunoblot analysis for Flag and NEK7 are shown. C LPS-primed BMDMs were treated with epalrestat, ponalrestat, ranirestat, and tolrestat for 1 h and then stimulated with ATP. Immunoblot analysis was used to detect cleaved caspase-1 and IL-1β in the Sup and the expression of NLRP3, caspase-1 p45, pro-IL-1β, and ASC in the Lys. D-E ELISA of IL-1β D and TNF-α E in the Sup was assessed from samples described in C. Data are presented as the mean ± SD from at least three biological samples. Statistical differences were analyzed using an unpaired Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001; ns not significant