Fig. 5

Both NLRP3/NF-κB inhibition and Nrf2 promotion are verified in effect of baicalein on MDSCs in vitro. MDSCs were treated with or without baicalein 3 h after being primed for 1 h for different concentrations (0.01 μM, 0.02 μM, 0.04 μM), then treated with or without ATP (5 mM) for another 30 min. a, b ROS and MitoSOX production in MDSCs was analyzed by flow cytometry. c, d IL-1β and IL-18 secretion was detected by ELISA. e Relative expression of Arg-1, P47 ^phox, GP91 ^phox, iNOS, NQO-1, and HO-1 mRNA in MDSCs were measured by QPCR. f Nuclear levels of Nrf2 and the protein expression of HO-1 were determined by western blot. g Immunofluorescence of Nrf2. h Western blotting was used to assess the expression levels of NLRP3, Pro-casp-1, and Pro-IL-1β proteins in the cell lysates, and Casp-1-p20 and mIL-1β in culture supernatants, respectively; GAPDH was used as a loading control for cell lysates. i P-NF-κB and NF-κB were determined by western blot. BA represents baicalein. Data represent the mean scores ± SEM of triplicate experiments. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ^#P < 0.05, and ^##P < 0.01 vs LPS with the ATP group