Figure 2

Short range RT-PCR assays for human SHBG gene expression. A. SHBG exon 5–8 assay. First strand cDNAs were generated from MCF-7, MCF-7-Tr LNCaP, HepG2 cells and normal human testis tissue using an oligo dT primer (see methods). Three RT-PCR transcripts were generated in the MCF-7 and LNCaP lanes, and four in the HepG2 and testis lanes. MCF-7-Tr is an MCF-7 clone stably transfected with a plasmid constitutively expressing large amounts of SHBG_L. All bands were reamplified and sequenced. SHBG RT-PCR transcript structures are given on the right. Note- this is a merged figure- the DNA marker and transfection control lanes were consolidated from a different part of the gel. B. SHBG exon 2–8 assay. PCR primers, specific for exon 2 and exon 8, were used to amplify cDNAs prepared from HepG2, LNCaP, MCF-7, MCF-7-Tr, and normal human testis, liver, prostate, breast, and brain tissues. All bands from HepG2, LNCaP, MCF-7, and testis were reamplified and sequenced. Bands labeled "*" produced sequences that were inconsistent with the sizes of the transcripts, and which were identical to other characterized sequences. DNA marker sizes, in base pairs, are given on the left. RT-PCR fragment transcript structures and sizes are given on the right. Note- this is a merged figure- the transfection control lane was taken from a different part of the gel.