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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:25702*  Cite 
Description

MTLn3 cell cotransfected with FAK siRNA (target A) and two tandem Src SH2 phosphotyrosine-binding domains (YFP-dSH2), which specifically detects tyrosine phosphorylation at focal adhesions. CIL:25703 is the corresponding control experiment. Fluorescence imaging was performed using a 60×/1.40 oil objective on an inverted microscope (1X-70;Olympus) in a 37°C closed system as previously described. Glass-bottomed dishes (35 mm) were coated with 10 µg/ml fibronectin for 1 h at 37°C. Cells were plated in Ham’s F12 containing 5% FBS and 20 mM Hepes, pH 7.2, and were allowed to adhere for 3 h. Fluorescent images were collected using a cooled CCD camera (CoolSNAP FX; Hamamatsu Photonics) and captured into MetaVue every 1 min for 1 h. Time-lapse sequences from live fluorescence imaging were first subjected to high-pass filtration based on the water algorithm (Zamir et al., 1999) to remove diffuse background fluorescence. Movie corresponds to video 5 from J Cell Biol. 2009 Apr 20;185(2):357-70. Epub 2009 Apr 13.

Biological Sources
NCBI Organism Classification
Rattus
Cell Type
mammary adenocarcinoma
Cell Line
MTLn3
Cellular Component
focal adhesion
Biological Context
Biological Process
tyrosine phosporylation
FAK knocksown siRNA
Attribution
Names
Keefe T. Chan
Christa L. Cortesi
Anna Huttenlocher
Published
J Cell Biol. 2009 Apr 20;185(2):357-70. Epub 2009 Apr 13.
Pubmed
19364917
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL25702
Archival Resource Key (ARK)
ark:/b7295/w9cil25702
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EYFP
Processing History
watershed filtering
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 326px 0.24µm
Y 324px 0.24µm
Time 60 seconds 60