TLRs-triggered immune response was enhanced in <i>Gpr108</i>-null cells.

Abstract

(A) Primary macrophage cells derived from litter mated Gp108+/+ and Gpr108-/- mice were treated with LPS (1 μg/ml, TLR4), Poly (I:C) (5 μg/ml, TLR3), immiquimod (50 μg/ml, TLR7) and R484 (10 μg/ml, TLR7). IL-6 and TNFα production were measured by ELISA after 18 hours treatment. The expression of IFNβ was determined by RT-qPCR after 8 hours treatment. (B) IL-6 and TNFα production were measured in immortalized Gpr108+/+ and Gpr108-/- macrophages treated with LPS (100 ng/ml) at different time points. (C) IL-6 and TNFα production were measured in primary Gpr108+/+ and Gpr108-/- MEF cells treated with LPS (100 ng/ml) for 18 hours. (D) iBMDM cells were transduced with a lenti-virus NF-κB reporter containing luciferase and treated with LPS (100 ng/ml) and immiquimod (50 μg/ml) for 18 hours. The luciferase activities were measured as fold increase by normalizing to the untreated cells as 1. (E) BMDM cells were stimulated with LPS (100 ng/ml), Poly (I:C) (5 μg/ml), R484 (10 μg/ml) and CPG (1μM) at different time points. The phosphorylated IRF3, IKBα and phosphorylated IKBα were measured by immunoblotting analysis with actin protein as its loading control. Data were shown as mean±SD (A-D). *p < 0.05, **p < 0.01, ***p<0.001. All experiments were repeated for at least three times.</p