Figure 10

Effects of protease inhibitors on Giardia lamblia protease activities and Giardia-promoted NF-κB p65 protein degradation. (A) Zymograms of Giardia lysate 10 µg (C; control of proteolytic activity) with protease inhibitors EDTA (5 mM), PHEN (10 mM), E-64 (10 μM), LEUP (50 µM), PEPA (1 μM), PEPAP (1 µM), PMSF (1 mM), and AEBSF (1 mM). Molecular mass markers are shown on the left in kilodaltons (kDa). (B) LPS-activated macrophage-like cells lysates were incubated with G. lamblia extracts (10 µg) and with protease inhibitors EDTA, PHEN, E-64 and LEUP, PEPA, PEPAP, PMSF and AEBSF. NF-κB p65 subunit levels were analyzed by Western blot using a specific anti-p65 antibody and anti-tubulin antibody. The blot shown is representative of three blots yielding similar results. Each value represents the mean ± SEM from at least 3 experiments (^****p < 0.0001, LPS-activated macrophage-like cells versus Giardia treated extracts; ^###p < 0.001, ^####p < 0.0001, inhibitors versus Giardia; ns, not significant). Full-length blots are presented in Supplementary Fig. S11.