Figure 5

Identification and validation of Smad4-dependent TGF-β target genes. a) Smad4-dependent TGF-β target genes in 28-8 cells were identified through hybridisation of Clontech Cancer II arrays. At least three hybridisations were performed for each clone and each set of conditions. Shown as examples are sections of filters containing the probes for fibronectin and BigH3. The induction of PAI-1 was detectable but less clear because a very strong neighbouring signal faded over the PAI-1 locus (data not shown). b) Corresponding filters to those shown in figure 4 were hybridised with probes for BigH3, fibronectin and PAI-1. c) Smad4-levels were titrated in clone 28-8 through dilution of doxycycline. Northern blotting with the same probes as used in (b). d) TGF-β induction of 28-8 cells was repeated in the presence of cycloheximide