Fig. 7.

ANT1 knockdown suppresses activation of NLRP3 inflammasome. Two groups of THP-1-derived macrophages, i.e., with shRNA-Ctrl or shRNA-ANT1, were primed with 100 ng ml⁻¹ LPS for 3 h, and then stimulated with ATP (5 mM, 1 h), MSU (500 µg ml⁻¹, 2 h), or Nigericin (10 µM, 2 h), respectively. a, b ELISA of IL-1β and IL-18 in the culture supernatant. c Immunoblot analysis of cell lysates from THP-1-derived macrophages. d Immunoblot analysis of Co-IP from THP-1-derived macrophages. e, f Flow cytometry analysis of mitochondrial membrane potential by JC-1 staining e or mitochondrial ROS by MitoSOX staining f from THP-1-derived macrophages with shRNA-Ctrl or shRNA-ANT1 lentivirus, followed by LPS treatment and ATP or Nigericin stimulation. g Quantitative real-time PCR analysis of mtDNA released from ANT1-knockdown THP-1-derived macrophages and left unstimulated (medium) or primed with LPS and stimulated with ATP, MSU or Nigericin. *P < 0.05, **P < 0.01, one-way ANOVA for multiple comparisons. Data are representative of three independent experiments (mean and SEM of three independent samples in a, b, e–g)