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Fig. 7. Effect of TGF-
-induced ET-1 on endothelial cell migration. Transwell assays: BAEC were seeded on Transwell inserts and cell-migration capacity analyzed as described in the Materials and Methods. (A) Schematic representation of filter device and representative pictures taken at corresponding sides of the filter membrane showing non-migrating cells (upper surface) and migrating cells (lower surface) for the indicated experimental conditions (basal, TGF- alone, TGF- plus SB-431542 and TGF- plus bosentan). (B) Cell-migration capacity is shown in the bar graph as the ratio of migrating versus non-migrating cells expressed as a percentage of control (untreated cells) (mean ± s.d., n=3, *P<0.05 versus basal, #P<0.05 versus TGF- alone). The potential effect of bosentan on TGF- signaling was analyzed by transfection of ALK5- and ALK1-specific reporters under overexpression of ALK5 constitutively active (ca) (C) and ALK1 ca (D) forms in cells treated with medium alone or with 10 µM bosentan. Promoter activity was estimated by luminometry and values expressed as fold induction with respect to the corresponding control without activation (mean ± s.d., n=3, *P<0.05 versus control without activation).
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