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

CIL:13880*  Cite 

Tem1-CAAX (green) is targeted away from spindle pole bodies in wild-type cells (this image) but can also still localize to the SPB (CIL# 13881). The CAAX motif from Ras2 targets proteins to the cytoplasmic side of the membrane. tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-TEM1-CAAX from a CEN plasmid were grown on 2% raffinose/2% galactose and transferred to 2% glucose medium prior to image capture. A differential interference contrast (DIC) image is also shown. 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 S5D, top panels, in J Cell Biol. (2011) 192: 599-614. Images in Fig S5 include CIL # 13880, 13881.

Technical Details

Cells (MATa tem1::GAL-UPL-TEM1-TRP1 pRS316::eGFP-TEM1-CAAX) 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
Cellular Component
spindle pole body
Mauricio Valerio-Santiago
Fernando Monje-Casas
J Cell Biol. (2011) 192: 599-614.
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
widefield illumination
fluorescence microscopy
differential interference contrast microscopy
Parameters Imaged
fluorescence emission
optical path length gradient
Source of Contrast
distribution of a specific protein
boundaries between regions with different refractive index
Visualization Methods
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
formaldehyde fixed tissue
ethanol fixed tissue
Relation To Intact Cell
whole mounted tissue
Spatial Axis Image Size Pixel Size
X 218px 0.0642µm
Y 218px 0.0642µm