Zymographic analysis of media conditioned by Raji, Jurkat, and NC-37 cell lines stimulated with cytokines and the in vitro invasion assay. (A) Zymograms of media conditioned by cells incubated for 16 hours at 37°C and 5% CO₂ in serum-free IMDM in the absence (control) or presence of a cytokine. Zymography was performed using 10% polyacrylamide gels containing 2 mg/mL gelatin as described in Materials and Methods. The final concentrations of cytokines were 10 ng/mL TNF-, 100 ng/mL IL-6, and 100 ng/mL IL-10. Media conditioned by BHK and KG-1 cells were used as standards showing the positions of the 92 kD (MMP-9) and 72 kD (MMP-2) activities in the gels. (B) Matrigel invasion by Raji and Jurkat cells after stimulation with IL-6 and IL-10 or without (control) as described in Materials and Methods. The asterisks * indicate statistically significant differences in the percentage of migration (P = .046 for Raji and P= .0002 for Jurkat) in the presence of IL-6 versus control (without IL-6). (C) Effect of MMP inhibitors on the percentage of migration of IL-6–stimulated Raji and Jurkat cells. The final concentrations of anti–MMP-9 and anti–MMP-2 antibodies and recombinant TIMP-1, as well as the preincubation conditions are described in Materials and Methods. The basal migration of Raji and Jurkat cells was set at 100% (control) and the percentages of migration in the presence of the different inhibitors are represented as mean ± standard deviation relative to the control.