Fig. 6. Analysis of GDNF-CAT reporter genes in transfected cells. (A) Schematic of the GDNF genomic locus. Boxes represents exonic sequences; the black vertical bars represent prospective Pax2-binding sites. The reporter plasmid p2.4-CAT contains GDNF sequences extending from unique BamHI site in exon1 to HindIII site approximately 2.4 kb upstream, fused to the CAT gene. This corresponds to position -1260 to +1052, according to previous numbering (Tanaka et al., 2000). Plasmid p
Apa-CAT has a deletion of 250 bp within the 5' UTR that encompasses the Pax2-binding site PBS2. Plasmid pPBS2-CAT has a small 34 bp deletion around the Pax2-binding site (PBS2). (B) CAT activity of reporter plasmids in transfected NIH 3T3 cells. Reporter plasmids were transfected with increasing amounts of Pax2b and Pax2a expression plasmids. Fold activation was calculated relative to the basal level of reporter gene expression. Data are presented as average values for activation and error bars reflect one s.e.m.
