Figure 6

p38 MAPK and NFκB signaling pathways are involved in resistin-induced SDF-1 promoter activity. (A) SDF-1-1010-Luc activity was demonstrated in TSGH 9201 cells pretreated with vehicle (DMSO) and SB203580 (SB) individually 1 h, and then stimulated with 25 ng/mL resistin for 1 h. (B) NFκB p50 activation was determined by TF ELISA assays in TSGH 9201 cells pretreated with vehicle (DMSO) and SB203580 (SB) individually or in combination for 1 h, and then treated with resistin for 1 h. All bar graphs represent folds of CL in TSGH 9201 cells, mean ± SEM. *P < 0.05 versus CL. #P < 0.05 vs. vehicle control (DMSO). (C) NFκB p50 binding to SDF-1 promoter in TSGH 9201 cells after 1 h resistin stimulation was determined by ChIP assay. (D) TSGH 9201 cells were incubated with the p38 inhibitor SB 203580 (SB, 10 μM) for 1 h. Next, the cells were treated with 25 ng/mL resistin for 12 h. Effect of resistin on the phosphorylation of IκBβ as well as on p50 nuclear translocation. Nuclear and cytoplasm cell lysate proteins were prepared and analyzed by western blot.