HeLa cells transfection with siRNA for gene silencing using Lullaby

siRNA were transfected by Lullaby siRNA transfection reagent in HeLa cell line.

this article demonstrate the high efficiency of Lullaby, siRNA transfection reagent from OZ Biosciences to induce gene silencing in MDCK

article reference: J Cell Sci. 2013 Apr 2

ERK2 but not ERK1 mediates HGF-induced motility in non small cell lung carcinoma cell lines.

Radtke S, Milanovic M, Rossé C, de Rycker M, Lachmann S, Hibbert A, Kermorgant S, Parker PJ.

Abstract

Aberrant signalling of receptor tyrosine kinases (RTKs), such as c-Met, the receptor for hepatocyte growth factor (HGF), has been implicated in the oncogenesis of various tumours including non-small cell lung carcinoma (NSCLC). Through its pro-migratory properties, c-Met has been implicated specifically in the process of tumour metastasis demanding a better understanding of the underlying signalling pathways. Various players downstream of c-Met have been well characterised, including the extracellular-signal-regulated kinases (ERKs) 1/2. In a small interfering (si) RNA based high throughput wound healing screen performed in A549 lung carcinoma cells, we identified ERK2 but not ERK1 as a strong mediator of HGF-induced motility. This finding was confirmed in several NSCLC cell lines as well as HeLa cells. One known substrate for ERK kinases in cell migration, the focal adhesion protein paxillin, was also one of the hits identified in the screen. We demonstrate that HGF stimulation results in a time dependent phosphorylation of paxillin on serine 126, a process which can be blocked by inhibition of the ERK1/2 upstream kinase Mitogen-Activated Protein Kinase/ERK Kinase 1 (MEK1) or inhibition of glycogen synthase kinase (GSK) 3. Further we show that paxillin turnover at focal adhesions is increased upon HGF-stimulation, an effect that is dependent on serines 126 (GSK3 site) and 130 (ERK site) within paxillin. In line with the isoform specific requirement of ERK2 for HGF-mediated migration in lung tumour cell models, ERK2 but not ERK1 is shown to be responsible for paxillin S126 phosphorylation and its increased turnover at focal adhesions.

Lullaby^® is the ideal siRNA transfection reagent for gene silencing reaching up to 90% gene silencing. It protects siRNA from extracellular degradation and maintains high viability due to its bio-degradable properties.