Figure 1

RasGrf1 expression in pancreatic islets. (A) RT-PCR amplification of the RasGrf1 locus. Total RNA samples from pancreatic islet preparations isolated from RasGrf1 knockout (KO) or wild type (WT) mice were tested by means of RT-PCR assays using oligonucleotides LM5 and LM85 (described in Methods) designed to amplify a specific 276 bp fragment corresponding to the catalytic domain of RasGrf1. C: negative control. MWM: molecular weight markers. (B) Analysis of RasGrf1/2 protein expression. Total protein extracts from WT and RasGrf1 KO pancreatic islets were immunoprecipitated with anti-RasGrf1/2 antibody and later submitted to SDS-PAGE followed by Western immunoblot with the same antibody (see Methods). The amount of protein in each lane was controlled and normalized by means of Western immunoblots using specific alpha-actin antibodies (lower panel).