Technical Reference #752

752.

Ca2 Influx Is An Essential Component Of The Positive-Feedback Loop That Maintains Leading-Edge Structure And Activity In Macrophages John H. Evans* and Joseph J. Falke, University of Colorado, PNAS, 104(752), (2007)
Link To Paper

Abstract:
In migrating eukaryotic cells phosphatidylinositol 3-kinase (PI3K) filamentous actin (F-actin) and monomeric Rho GTPases are keycomponents of a complex positive-feedback system that maintainsand amplifies a phosphatidylinositol-345-trisphosphate

Keywords:
actin calcium chemotaxis phosphoinositide 3-kinase phosphatidylinositol-3;4;5-trisphosphate

Materials & Methods:
Before experiments stably transfected cells were cultured on glassbottomed 35-mm dishes (MatTek Ashland MA) in culture medium without antibiotics at 1 105 cells per plate for a minimum of 3–4 h to allow cells to establish a polarized morphology.

Microscopic Technique
Fluorescence Microscopy

Cell Type(s)
RAW264.7