Requirement of MAP kinase for differentiation of fibroblasts to adipocytes, for insulin activation of p90 S6 kinase and for insulin or serum stimulation of DNA synthesis.

摘要: A phosphorothioate-oligonucleotide-based antisense strategy for depleting MAP kinase was developed. The 17mer probe, EAS 1, caused a potent and concentration-dependent decrease in the steady state expression of p42 p44 3T3 L1 fibroblasts adipocytes with submicromolar concentrations effective. Antisense 1 elicited dose-dependent inhibition insulin- serum-stimulated DNA synthesis. Elimination by > 95% to levels undetected blocked ability serum insulin stimulate synthesis 87-95%. differentiation into prevented microM 1. corresponding sense, scrambled or sense plus phosphorothioate oligonucleotides did not deplete from either cell type, inhibit stimulation interfere differentiation. Two kinases on different activation pathways were depleted whereas activate p90 S6 90% eliminated 4.5 In conclusion these results show that is required synthesis, activity cells. Moreover, development probe against target sequence conserved across range species provides molecular tool general applicability further dissecting precise targets roles kinase.