Fig. 1.

5’ Genomic organisation of Fgf-10. A novel Fgf-10 transcription start site 653 nucleotides 5’ to the Entrez published site was identified by RLM-RACE using total RNA isolated from embryonic day 18.5 mouse lung (A). Analysis of the potential promoter region of Fgf-10 (−500 to +1 bp) identified a string of 12 PEA3-binding sites approximately 360 to 240 bp upstream of the novel start site. The novel FGF-10 cDNA was confirmed by PCR analysis of cDNA and genomic (gDNA) samples (from E18.5 mouse lung) using forward primers specific to the genomic sequence immediately upstream of the predicted start site (grey) and at the predicted start of exon 1 (black), together with a common reverse primer (black) (B). PCR products of the expected size were obtained from both cDNA and gDNA samples when using the inner primer pair, confirming the predicted Fgf-10 start site was transcribed. The gDNA primer set yielded no product with the cDNA sample, as predicted, but amplified a clear band in the gDNA sample. Primers specific for cDNA (housekeeping gene hPRT, with primers spanning an intron to prevent amplification of gDNA) and gDNA (known intron sequence from Fgf-3) confirmed the integrity of both DNA samples and the lack of gDNA contamination.