FIGURE 2.

FnIII-1c activates the NFκB signaling pathway in human dermal fibroblasts. A, monolayers of human fibroblasts were serum-starved overnight and then treated with the designated amounts of FnIII-1c, FnIII-13, or FnIII-10n in 0.1% BSA/DMEM either for 1 h or for the designated time. Control cells (0) received PBS. The nuclear fraction was isolated and analyzed by Western blot for the presence of the NFκB protein p65/rel A. The membranes were then stripped and reprobed with an antibody against nuclear lamin A/C as loading control. B–D, the cytosolic fraction (B and C) or the total cell lysate (D) was electrophoresed and immunoblotted using antibodies against phosphorylated IKK (p-IKK) (B), phosphorylated IκBα (p-IκBα), or total IκBα (IκBα) (C and D). The membranes were then stripped and reprobed with antibodies against FAK or β-actin as loading control. Blots shown are representative of one experiment performed on three separate occasions.