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

CIL:32024*  Cite 
Description

Video shows the dynamics of microtubules (MTs) using a GFP fusion construct of EB3 (end binding protein 3) that transiently binds to growing MT plus ends, generating a punctate pattern of EB3-GFP comets throughout the cell. This video shows the number of EB3-GFP comets detected after nocodazole perturbation in an RCC-4 cell that is VHL -/- (Von Hippel-Lindau). Compare with experimental video (RCC-4 supplemented with active VHL) in CIL # 26571; there is an increased number of growing MTs when VHL is active. RCC-4 cells were infected with a retroviral vector to express EB3-GFP. Cells on round coverslips were transferred to a homemade holding device, and 600 µl of the appropriate medium, containing 40 nM nocodazole was added. The video was acquired with a microscope (IX70 Delta Vision Spectris; Olympus), temperature-controlled at 37C, using a 60× NA 1.4 differential interference contrast (DIC) oil Plan-Apochromat objective, ex 470 em 520, and a camera (CoolSNAP HQ; Roper Industries) with an exposure time of 100 ms and a frame rate of 0.5 s. Acquisition software used was SoftWoRx version 3.3.4 (Applied Precision). This is the raw image file for the left half of Video 5 in J Cell Biol. 2010. 190: 991-1003.

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
renal cell carcinoma
epithelial cell
Cell Line
RCC4
Cellular Component
microtubule
cytoskeleton
microtubule plus end
Biological Context
Biological Process
protein polymerization
Molecular Function
protein binding
Attribution
Names
Claudio R. Thoma
Alexandre Matov
Katrin L. Gutbrodt
Christian R. Hoerner
Zlatko Smole
Wilhelm Krek
Gaudenz Danuser
Published
J Cell Biol. 2010. 190: 991-1003.
Pubmed
20855504
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL32024
Archival Resource Key (ARK)
ark:/b7295/w9cil32024
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
widefield illumination
time lapse microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 768px 0.1103µm
Y 768px 0.1103µm
Time 0.5 seconds 80