Figure 3

Effect of the TLR4 antagonist in blocking resistin-induced expression of SDF-1 and p38 MAPK. (A) TSGH 9201 cells were kept as control (CL) or stimulated with 25 ng/mL resistin. Cells were pretreated with or without 1 or 2 mg/ml LPS-RS for 1 h and then stimulated with 25 ng/mL resistin for 4 h. SDF-1 mRNA levels were determined through real-time PCR in TSGH cells and normalized to 18S rRNA. (B) SDF-1-1010-Luc activity were determined in TSGH cells with or without 1 or 2 mg/ml LPS-RS for 1 h. (C) TSGH cells were kept as CL or stimulated with resistin for 30 min, and the phosphorylation of p38 was determined by western blotting. All bar graphs represent folds of CL cells. *P < 0.05 versus CL. #P < 0.05 versus vehicle control (DMSO). (D) TSGH 9201 cells were transfected with control siRNA (si-CL) or a specific siRNA of si-TLR4 and the cells were treated with 25 ng/mL resistin for 30 min and 6 h, and then the phosphorylation of p38 and TLR4 were determined by western blotting. Human SDF-1 secretion was determined by ELISA.