Figure 4
From: Secreted Thrombospondin-1 Regulates Macrophage Interleukin-1β Production and Activation through CD47
In human cells TSP1 inhibits IL-1β and caspase-1 mRNA, but not NLRP3 induced in the presence of LPS.
(A–C) Differentiated THP-1 cells were cultured with soluble human TSP1 and after 15–20 min were either left untreated or stimulated with 20 ng/ml LPS for an additional 3 h. (D–F) Primary human macrophages were treated with TSP1 (as shown) or OptiMEM alone for 15–20 min and then 20 ng/ml LPS was added for 3 h. (A,D) IL-1β, (B,E) NLRP3 and (C,F) caspase-1 mRNA expression was analyzed by quantitative real-time PCR. (G) Bone marrow from wild type and cd47−/− mice was taken and differentiated into BMDMs. Cells were pre-treated with of recombinant domains of TSP1 (22 pM of E123CaG1 or 3TSR) or OptiMEM alone for 15–20 min and then 20 ng/ml LPS or medium alone was added for 3 h. The level of IL-1β mRNA quantified using real-time PCR. (H) Recombinant domains of TSP1 and their relevant receptor specificities. For panels A through G significance was calculated using one-way parametric ANOVA. When data significance is indicated “*” means p < 0.05, “**” means p < 0.01 and “***” means p < 0.005. Data shown is a representative of 3 independent experiments performed.
