Figure 5.

Regulation of synovial fibroblast production of IL-6 and CCL5 via distinct signaling pathways. Synovial fibroblasts from patients with RA were seeded and treated with either DMSO carrier control or the specified pathway inhibitors for 1 hour. All wells were then treated with recombinant galectin 3 (10 μg/ml). A and B, Production of IL-6 (A) and CCL5 (B) after 24 hours of stimulation, as analyzed by ELISA. Bars show the mean and SD of duplicate assays combined from at least 3 patients. C–F, Western blots of cell lysates of synovial fibroblasts obtained from patients with RA and stimulated with galectin 3 for the indicated time periods. Cell lysates were labeled using primary antibodies to ERK (C), JNK (D), p38 (E), and Akt (F) and their active phosphorylated forms. Results are representative of 4 lines or are from 4 lines combined. G, Binding of the activated p65 NF-κB subunit to an NF-κB consensus sequence in nuclear extracts from RA synovial fibroblasts stimulated with medium (control), TNFα (10 ng/ml), or galectin 3 (10 μg/ml), as analyzed using a TransAm NF-κB ELISA kit. Bars show the mean and SD. * = P < 0.05; ** = P < 0.01; *** = P < 0.001, versus DMSO or control medium. Bis = bisindolylmaleimide I; NS = not significant (see Figure 4 for other definitions).