Mouse Embryonic Fibroblasts (MEF) were efficiently transfected using DreamFect Gold transfection reagent.

Mouse Embryonic Fibroblasts (MEF) cultured in 35mm pre-coated with 0.1% collagen were transfected in suspension with 1.6µg plasmid using DreamFect™Gold

This article demonstrates that due to its low toxicity level and to its high transfection capacities, DreamFect Gold from OZ Biosciences is the reagent of choice for transfecting Mouse Embryonic Fibroblasts.

article reference: J Biol Chem. 2013 Nov 21.

β-arrestin promotes Wnt-induced Lrp6 phosphorylation via increased membrane recruitment of Amer1.

Kriz V, Pospichalova V, Masek J, Kilander MB, Slavik J, Tanneberger K, Schulte G, Machala M, Kozubik A, Behrens J, Bryja V.

Abstract

β-arrestin is a scaffold protein, which regulates signal transduction by seven transmembrane spanning receptors. Among other functions it is also critically required for Wnt/β-catenin signal transduction. In the present study we provide for the first time a mechanistic basis for the β-arrestin function in Wnt/β-catenin signaling. We demonstrate that β-arrestin is required for efficient Wnt3a-induced Lrp6 phosphorylation, a key event in downstream signaling. β-arrestin regulates Lrp6 phosphorylation via a novel interaction with phosphatidylinositol (4,5) bisphosphate (PtdIns(4,5)P2)-binding protein Amer1/WTX/Fam123b. Amer1 has been shown very recently to bridge Wnt-induced and Dishevelled (Dvl)-associated PtdIns(4,5)P2 production to the phosphorylation of Lrp6. Using FRAP we show here that β-arrestin is required for the Wnt3a-induced Amer1 membrane dynamics and downstream signaling. Finally we show that β-arrestin interacts with PtdIns kinases PI4KIIα and PIP5KIβ. Importantly, cells lacking β-arrestin showed higher steady state levels of the relevant PtdInsP and were unable to increase levels of these PtdInsP in response to Wnt3a. In summary, our data show that β-arrestins regulate Wnt3a-induced Lrp6 phosphorylation by the regulation of the membrane dynamics of Amer1. We propose that β-arrestins via their scaffolding function facilitate Amer1 interaction with PtdIns(4,5)P2, which is produced locally upon Wnt3a stimulation by β-arrestin- and Dvl-associated kinases.

DreamFect™Gold allows transfecting all types of nucleic acids with a very high efficiency. Due to its formulation, DreamFect™Gold delivers a large quantity of nucleic acids leading to higher protein expression compared to other transfection reagents. It is fully biodegradable and does not interfere with cellular mechanisms.