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

CIL:13882*  Cite 
Description

Tem1 normally localizes preferentially to the spindle pole body (SPB) that enters the daughter cell during anaphase (this image and CIL# 13885). This image shows SPB localization of eGFP-Tem1 (eGFP, green) in metaphase cells. tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-TEM1 from a CEN plasmid were grown on 2% raffinose/2% galactose and transferred to 2% glucose medium prior to image capture. Nuclear morphology was assessed by DAPI (blue). A differential interference contrast (DIC) image is also shown (gray). The tem1Δ::GAL1-UPL-TEM1 strain allows for the rapid, conditional depletion of Tem1. UPL, which stands for ubiquitin-proline-LacI, acts as a destabilizing module that permits rapid degradation of appended proteins. Image is Fig 1A, top panels, in J Cell Biol. (2011) 192: 599-614. Other images in Fig 1 include CIL #13882, 13883, 13884, 13885, 13886, 13887.

Technical Details

Cells (MATa tem1::GAL-UPL-TEM1-TRP1 pRS316::eGFP-TEM1) were fixed in 2.5% formaldehyde for 10 min, washed twice, and resuspended in 0.1 M potassium phosphate buffer, pH 6.4. Cells were then fixed for 10 min in 80% ethanol and resuspended in 1 mg/ml DAPI. Imaging was performed at 25C using a Leica DM6000 microscope equipped with a 100x/1.40 NA oil immersion objective lens, A4, L5, and TX2 filters, and a digital CCD camera (DFC350, Leica). Pictures were processed with LAS AF (Leica) and ImageJ software.

Biological Sources
NCBI Organism Classification
Saccharomyces cerevisiae
Cell Line
W303
Cellular Component
nucleus
spindle pole body
Attribution
Names
Mauricio Valerio-Santiago
Fernando Monje-Casas
Published
J Cell Biol. (2011) 192: 599-614.
Pubmed
21321099
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13882
Archival Resource Key (ARK)
ark:/b7295/w9cil13882
Grouping This image is part of a group.
Sample Preparation
Methods
formaldehyde fixed tissue
ethanol fixed tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 218px 0.0642µm
Y 218px 0.0642µm