Ki16425 attenuates LPA1 / CD14 association and LPS-induced signaling and cytokine release. A. Cell lysates from CD14-V5 and LPA1-Myc over-expressed HEK293 cells were incubated with DMSO or ki16425 (5 µM) for 1 h. Cell lysates were then subjected to immunoprecipitation with a Myc tag antibody, followed by a V5 tag immunoblotting. Input lysates were analyzed with V5 tag, Myc tag, and β-actin immunoblotting. B. HepG2 cells were treated with ki16425 prior to LPS (1 µg/ml, 30 min) treatment. Cell lysates were analyzed by phospho (P)-PKCδ, P-p38 MAPK, and β-actin immunoblotting. C. Raw264 cells were treated with DMSO or Ki16425 (5 µM) for 30 min prior to LPS treatment (0.2 µg/ml, 3 h). IL-6 release in medium was measured by ELISA.