FIGURE 6.

Pro-IL-1β and GPSM3 are reciprocally regulated during LPS-induced inflammasome priming. BMDMs were generated from WT or Gpsm3(−/−) mice and primed with LPS for 3 h with 100 ng/ml and/or subsequently treated with HLA for 1 h, as indicated. Il1b and Nlrp3 mRNA levels in each cell line were assessed using quantitative RT-PCR as described under “Materials and Methods” (panels A and B). THP1 cells were treated with LPS (100 ng/ml) or vehicle for 3 h. GPSM3, IL1B, and NLRP3 mRNA levels were assessed by quantitative RT-PCR (panels C, D, and E). Data are expressed relative to the β2 M gene transcript; *, p < 0.05 by one-way ANOVA. GPSM3 and actin (loading control) protein levels were also assessed using immunoblot analysis with the indicated antibodies (panel F). Samples from three independent experiments are shown.