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. 2014 May 29;111(1):139–148. doi: 10.1038/bjc.2014.239

Figure 1.

Figure 1

EDPs influence on Hsp90, pro-MMP-2 and uPA secretion. (A) Gelatin zymography, gelatin plasminogen zymography and western blotting analysis of conditioned media and actin content in total cell extracts from HT-1080 cells cultured for 6, 24 and 48 h in the absence (−) or presence of 50 μg ml−1 Kel (+) as described in Materials and Methods section. (B) Quantitative evaluation of pro-MMP-2 secretion by HT-1080 cells on gelatin zymography. Results (mean±s.e.m.) were expressed as the percentage of control 6 h (EDP-untreated cells). *P<0.005, **P<0.001, significantly different from control. Lighter bars, untreated cells; black bars, EDP-treated cells. (C) Quantitative evaluation of uPA secretion by HT-1080 cells on gelatin plasminogen zymography. Results (mean±s.e.m.) were expressed as the percentage of control for 6 h (EDP-untreated cells). *P<0.005, **P<0.001, significantly different from control. Lighter bars, untreated cells; black bars, EDP-treated cells. (D) Quantitative evaluation of Hsp90 secretion by HT-1080 cells by western blotting. Results (mean±s.e.m.) were expressed as the percentage of control for 6 h (EDP-untreated cells). **P<0.001, significantly different from control. Lighter bars, untreated cells; black bars, EDP-treated cells. (E) Dose-effect study of EDP treatment (Kel 0–100 μg ml−1) on pro-MMP-2, uPA and Hsp90 secretion. Gelatin zymography, gelatin plasminogen zymography and western blotting analysis of conditioned media and actin content in cell extracts from HT-1080 cells cultured for 24 h in the absence (Ctl) or in the presence of 1–100 μg ml−1 Kel. (F) Gelatin zymography, gelatin plasminogen zymography and western blotting analysis of conditioned media from HT-1080 cells cultured for 24 h in the absence (Ctl) or in the presence of 50 μg ml−1 Kel (Kel) or synthetic elastin peptides (VGVAPG, AGVPGLGVG, GRKRK). All synthetic peptides were used at 200 μg ml−1. Quantitative evaluation of protein secretions by HT-1080 cells on gelatin zymography, gelatin plasminogen zymography and by western blotting. Results (mean±s.e.m.) were expressed as the percentage of control (EDP-untreated cells).