Fig. 3
Epalrestat blocks ASC oligomerization but does not affect K+ or Ca2+ efflux. A Immunoblot analysis of BMDMs treated with epalrestat for 1 h before LPS stimulation for 4 h or BMDMs treated with epalrestat for 1 h after LPS stimulation for 1 h. B ELISA of TNF-α from samples described in A. C–D LPS-primed BMDMs were treated with epalrestat (40 μM) before stimulation with ATP, nigericin, SiO2, or poly (I: C) and Pam3CSK4-primed BMDMs were treated with epalrestat (40 μM) before stimulation with LPS. Immunoblot analysis of epalrestat was used to detect cross-linked ASC in the Triton X-insoluble pellet. E Quantification of intracellular potassium levels in LPS-primed BMDMs pre-treated with various doses of epalrestat and stimulated with nigericin. F A trace showing ATP-induced Ca2+ flux was measured using a FLIPRT Tetra system in LPS-primed BMDMs treated with epalrestat. 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