FIG 5.

Mechanisms involved in mediating the secretion of IL-1β by R. australis-infected macrophages. BMMs were isolated from B6, TLR4^−/−, MyD88^−/−, and ASC^−/− mice. BMMs were infected with R. australis at an MOI of 6 or stimulated with 50 ng/ml of purified R. australis LPS plus ATP for 24 h. ATP was added 1 h prior to collection. Supernatant was collected from infected BMMs of B6, TLR4^−/− (A), and MyD88^−/− (B) mice. Levels of secretion of IL-1β by these infected macrophages were evaluated by ELISA. (C) The synthesis of pro-IL-1β in BMMs of B6, ASC^−/− and TLR4^−/− mice was evaluated by specific antibodies against pro-IL-1β by immunoblotting after the cell lysates were collected. Salmonella LPS plus ATP served as the positive control. Data represent two independent experiments.