Figure 5
From: Secreted Thrombospondin-1 Regulates Macrophage Interleukin-1β Production and Activation through CD47
CD47 associates with CD14 and is dissociated by TSP1.
Differentiated THP-1 cells were incubated for 15 min in the presence of (A) 22 pM soluble human TSP1 or (C) 22 pM of recombinant 3TSR, NoC1, or E123CaG1. (B) Primary human monocyte derived macrophages were incubated with increasing concentrations of human soluble TSP1 (0.1, 1, 10, 100 ng/ml) for 15 min. CD14 was precipitated using or IgG-k anti-mouse isotype control antibody or mouse anti-human CD47 (B6H12) monoclonal antibody. Bound = protein that precipitated with respective antibodies; Unbound = lysate after precipitation by the beads. (D–E) THP1-XBlue cells were differentiated stimulated with indicated concentrations of TSP1 for 15–20 min. Cells were left untreated or (D) treated with 20 ng/ml LPS or (E) 100 ng/ml Pam3CSK4 for 3 h. Sterile QuantiBlue was added to quantify NFκB/AP-1 with the inducible reporter (SEAP) system and cells were incubated for an additional 2 h. In panels D and E significance was calculated using one-way parametric ANOVA. When data significance is indicated “**” means p < 0.01. ND = not detected. Data shown is a representative of 3 independent experiments performed.
