Figure 4.

EDP increases extracellular Hsp90 and promotes pro-MMP-2 and uPA accumulation in HT-1080 culture media. (A) Gelatin zymography and gelatin plasminogen zymography analysis of Hsp90 inhibitors effect (1.5 μM geldanamycin and 10 μM radicicol) on the production of pro-MMP-2 and uPA in the presence (+: 50 μg ml⁻¹) or in the absence (−) of EDPs (24 h of treatment). (B) Quantitative evaluation of protein secretions by HT-1080 cells on gelatin zymography and gelatin plasminogen zymography. Results (mean±s.e.m.) were expressed as the percentage of control (EDP-untreated cells). *P<0.005, **P<0.001, significantly different from Kel alone. (C) Gelatin zymography and gelatin plasminogen zymography analysis of monoclonal anti-Hsp90 antibody effect on the production of pro-MMP-2 and uPA in EDP-treated HT-1080 cells (50 μg ml⁻¹) (24 h of treatment). (D) Quantitative evaluation of protein secretions by HT-1080 cells on gelatin zymography and gelatin plasminogen zymography. Results (mean±s.e.m.) were expressed as the percentage of control (EDP-untreated cells). *P<0.005, **P<0.001, significantly different from Kel alone. (E) Gelatin zymography and gelatin plasminogen zymography analysis of the effect of recombinant Hsp90α protein on the production of pro-MMP-2 and uPA by HT-1080 cells (24 h of treatment). (F) Quantitative evaluation of protein secretions by HT-1080 cells on gelatin zymography and gelatin plasminogen zymography. Results (mean±s.e.m.) were expressed as the percentage of control (EDP-untreated cells). *P<0.005, **P<0.001, significantly different from control.