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

CIL:13883*  Cite 

Constitutive targeting of Tem1 to the spindle pole body (SPB) in metaphase cells (this image) and anaphase cells (CIL#13886). tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-CNM67–TEM1 from a CEN plasmid were grown on 2% raffinose/2% galactose. Image shows localization of Cnm67-Tem1 (eGFP, green) in metaphase after cells were transferred to medium with 2% glucose. Tem1 normally localizes preferentially to the SPB that enters the daughter cell during anaphase (CIL# 13882, 13885). Cnm67 is an integral component of the outer plaque of the SPB and the Cnm67-Tem1 fusion is found to be symmetric from SPB duplication until the end of mitosis. 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, middle 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-CNM67–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
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.
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