Identification of RE-1-containing genes potentially involved in beta cell survival.

Abstract

<p><i>A</i>. Alignment of identified RE-1 sequences in human (hum) or murine (mus) genes with the consensus RE-1. Left alignment identifies members of the insulin-like growth factor transduction pathway: <i>Igfbp7</i>: insulin-like growth factor binding protein 7; <i>IRS</i>: insulin receptor substrate; <i>Akt3</i>: PKB gamma; <i>CTNND2</i>: delta2 catenin; <i>Ctnnb1</i>: beta catenin; <i>MAPK10</i>: JNK3; <i>MAP4K3</i>: MEK kinase kinase 3. Upper right alignment identifies members of the cyclin-related family of mitogenic factors: <i>Cdk5r2</i>: cyclin-dependent kinase 5 regulatory subunit 2; <i>CKS2</i>: CDC28 protein kinase regulatory subunit 2; <i>CNNG1</i>: cyclin G1; <i>Cnnm2</i>: cyclin M2. Lower right alignment identifies <i>Api5</i>: apoptosis inhibitor 5 and <i>Nrg1</i>: neuregulin1. <i>B</i>. Quantitative RT-PCR analysis of mRNA levels from control INS-1E cells (white bars) and INS-1E cells infected with GFP-expressing adenovirus (gray bars) or <i>REST</i>-expressing adenovirus (black bars). <i>Ptprn</i>: protein tyrosine phosphatase, receptor type, N; <i>Irs2</i>: insulin receptor substrate2; <i>Cdk5r2</i>: cyclin-dependent kinase 5 regulatory subunit 2; <i>Nrg1</i>: neuregulin1; <i>Ctnnd2</i>: delta2 catenin; <i>Api5</i>: apoptosis inhibitor 5. Results are mean ± SD of six independent experiments. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 versus INS-1E cells transduced with GFP. <i>C</i>. Quantitative RT-PCR analysis of mRNA levels from islets of WT (white bars) (n = 5) and RIP-REST (black bars) (n = 5) mice. <i>Gjd2</i>: Connexin36; <i>Mapk8ip1</i>: Ib1; <i>Cdk5r2</i>: cyclin-dependent kinase 5 regulatory subunit 2; <i>Ptprn</i>: protein tyrosine phosphatase, receptor type, N; <i>Irs2</i>: insulin receptor substrate2; <i>Nrg1</i>: neuregulin1; <i>Ctnnd2</i>: delta2 catenin. Results are mean ± s.e.m. **<i>P</i><0.01 versus values of wild type animals. D. Left panel, immunoblotting of total proteins from non infected (NI) and infected (Ad-GFP, Ad-REST) INS-1E cells showing decreased protein levels of Cdk5r2 upon REST expression. The right panel shows the corresponding quantifications of REST protein levels in INS-1E cells after infection. Data are mean ± SEM of 4 independent experiments. **<i>P</i><0.01, ***<i>P</i><0.001 versus the respective Ad-GFP-infected and NI conditions.</p