Figure 3

A) Effect of AMSH-2 on TGF-β induced transcriptional activity of 3TP driven luciferase. HepG2 cells were cotransfected with 3TP-lux reporter together with the empty vector or increasing amounts of AMSH-2 Myc construct. Twenty-four hours posttransfection, cells were serum-starved and left untreated (white bars) or stimulated (black bars) with 5 ng/ml TGF-β1 for 16–20 h before the luciferase and β-galactosidase activities were measured. B) Effect of AMSH-2 on TGF-β induced transcriptional activity of PAI-1 luciferase. HepG2 cells were cotransfected with PAI-1 reporter together with the empty vector or 0.9 μg/ml of AMSH-2 Myc construct. A similar procedure as described in A was followed. C) AMSH-2 does not stimulate NF-kB signaling pathway. 293 cells were cotransfected with ELAM luciferase reporter construct together with an empty vector, 0.9 μg/ml of AMSH-2 Myc construct or 0.9 μg/ml of TRAF-2, used as a positive control. Forty-eight hours posttransfection, luciferase and β-galactosidase activities were measured.