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  • FAK-Src signalling is important to renal collecting duct morphogenesis: discovery using a hierarchical screening technique.

FAK-Src signalling is important to renal collecting duct morphogenesis: discovery using a hierarchical screening technique.

Biology open (2013-04-26)
Guangping Tai, Peter Hohenstein, Jamie A Davies
ABSTRACT

This report describes a hierarchical screening technique for identification of pathways that control the morphogenesis of the renal collecting duct system. The multi-step screen involves a first round using a 2-dimensional, cell-line-based scrape-healing assay, then a second round using a 3-dimensional tubulogenesis assay; both of these rounds use new cell lines described in this report. The final stage is ex vivo organ culture. We demonstrate the utility of the screen by using it to identify the FAK-Src-pathway signalling as being important for collecting duct development, specifically for the cell proliferation on which this development depends.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Y-27632 dihydrochloride, ≥98% (HPLC)
Sigma-Aldrich
Anti-Mouse IgG (whole molecule)–FITC antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
ITS+1 Liquid Media Supplement (100×), liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Anti-Rat IgG (whole molecule)–FITC antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-Rabbit IgG (whole molecule)–TRITC antibody produced in goat, affinity isolated antibody, buffered aqueous solution