Figure 5.

Effect of endotoxin tolerance on expression of Tollip, SOCS-1, IRAK-M, and SHIP-1. Human monocytes were pretreated for 20 h with medium or 10 ng/ml LPS, washed and treated with medium, or restimulated with 100 ng/ml LPS as shown. RNA was isolated, reverse-transcribed, and analyzed by real-time PCR to determine expression levels of Tollip, SOCS-1, IRAK-M (A), and SHIP-1 (B). The results of five independent experiments (mean ± sd) are shown. (C) and (D) Whole cell lysates were examined by Western blot analyses to determine protein expression of Tollip, IRAK-M (C) and SOCS-1 (D), using the corresponding Abs. Total p38 and β-actin immunoblots were run to control for protein loading, and IκB-α and p-p38 proteins were immunoblotted to monitor for LPS inducibility/tolerization. Results of a representative experiment (n=3) are shown.