Fig. 1

GTK-expression in transgenic islets. (A) Transcription from the transgene was studied by RT-PCR and Southern blotting on equal amounts of DNase treated RNA from 3-month-old mice using a 5′-primer matching nucleotide 1502–1522 of the GTK cDNA and a 3′-primer corresponding to the last 18 nucleotides of the myc-epitope. Samples not treated with reverse transcriptase (RT) were run in parallel. As control RT-PCR for β-actin was performed (top). One experiment was performed. (B) Isolated islets were cultured as above, immunoprecipitated with anti-GTK antibody, and subjected to Western blot analysis for GTK (top) or lysed in SDS sample buffer supplemented with protease inhibitor, and subjected to Western blot analysis for GTK (bottom). The result is based on four independent experiments.