Fig. 3.

EMSA indicates the specificity of the Smad3/4 binding sequence identified in the AChE promoter. a Electrophoretic mobility shift assay using the ³²P-labeled potential Smad binding sequence as a probe, incubated with nuclear extracts from HeLa cells treated with or without A23187 (2 μM) (lane 2–5) or TG (1 μM) (lane 6–9) for the indicated time. b ³²P-labeled probe was incubated with nuclear extracts from normal HeLa cells in the presence of oligonucleotide competitors: Wt Smad3/4 (lane 3), Mt Smad3/4 (lane 4), Wt YY1 (lane 5), Mt YY1 (lane 6), Wt NF-Y/CBF (lane 7), Mt NF-Y/CBF (lane 8) (Wt wild-type, Mt mutant-type). c Interference assays using Smad3/4 consensus oligonucleotides as competitor. The ³²P-labeled probe was incubated with nuclear extracts from normal (lane 1–3) or apoptotic HeLa cells treated by TG (1 μM) for 48 h (lane 4 and lane 6)