Figure 2. Inhibition of Lyp decreases TLR responses in monocytes.

CD14⁺ monocytes were isolated from freshly thawed PBMC and rested for 2 hours before addition of A) Lyp inhibitor I-C11 (5 μM) or B) 8b (1 μM) or DMSO control. After 30 minutes monocytes were treated with 0.1 ng/ml LPS (S. minnesota R595, List Biological Laboratories, Campbell, CA, USA) and supernatants were collected after 16 hours. TNF and IL-6 were measured by ELISA (eBioscience, San Diego, CA, USA). These inhibitors did not cause cytotoxicity during the overnight culture as measured by LDH release assay (J. Pottle and J.A. Hamerman, unpublished observations). Statistical significance was determined using a paired Wilcoxon matched pairs signed rank test.