1. The endogenous IGRP gene is selectively expressed in βTC-3 cells and not in Y1 cells. βTC-3 RNA (50 μg) and Y1 RNA (50 μg) were annealed to labeled oligonucleotide primers complementary to exon 1 of the IGRP gene (lanes 1 and 4), cyclophilin A gene (lanes 2 and 5), or both (lanes 3 and 6).
The endogenous IGRP gene is selectively expressed in βTC-3 cells and not in Y1 cells. βTC-3 RNA (50 μg) and Y1 RNA (50 μg) were annealed to labeled oligonucleotide primers complementary to exon 1 of the IGRP gene (lanes 1 and 4), cyclophilin A gene (lanes 2 and 5), or both (lanes 3 and 6). A primer extension assay was then performed (as described in research design and methods). A dideoxynucleotide sequencing reaction was also performed, and the products of this sequencing reaction (not shown) and the primer extension reaction were then separated on the same denaturing polyacrylamide gel. This allowed direct estimation of the sizes of the major extended IGRP and cyclophilin A (cyclo) products ([) and the calculation of their corresponding transcription start sites.
Larry J. Bischof et al. Diabetes 2001;50:
©2001 by American Diabetes Association