Fig. 3.

Inhibition of HMGCR impairs endotoxin tolerance. A, B: On day 0, peritoneal macrophages were seeded at 5 × 10⁶ cells per 60 mm dish. On day 1, cells were switched to medium containing (+) or not containing (−) 200 ng/ml LPSs or 25 μM compactin (CPN) as indicated (first treatment). After incubation for the indicated time, some cells were harvested for quantification of IL-1β mRNA through RT-QPCR, with the amount of the mRNA prior to the treatment set to one (A). The amount of IL-1β protein secreted into medium was determined through ELISA as well (B). On day 2, 24 h after the first treatment, the rest of the cells were switched to fresh medium containing the same concentration of LPSs and compactin as indicated (second treatment). After incubation for the indicated time, the amount of IL-1β mRNA inside the cells and that of IL-1β protein secreted into medium was determined as described above. Results are presented as mean ± SE from three independent experiments.