Valerio-Santiago, Mauricio; Monje-Casas, Fernando (2021). CIL:13865, Saccharomyces cerevisiae. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0R78CXC

3HA-Cdc14 (red) localization in anaphase. The protein is released throughout the whole cell. Anaphase is determined by spindle morphology (tubulin, green) and nuclear morphology (DAPI, blue). Image is Fig 3D, right panels, in J Cell Biol. (2011) 192: 599-614. Images in Fig 3 include CIL# 13862, 13863, 13864, 13865, 13866, 13867.

• 2021

• Monje-Casas, Fernando
• Valerio-Santiago, Mauricio

Cells (MATa cdc14::3HA-CDC14), grown in rich media with glucose, were fixed for 15 min in 3.7% formaldehyde and 0.1 M potassium phosphate buffer, pH 6.4. Cells were then washed twice with 0.1 M potassium phosphate buffer, pH 6.4, and resuspended in 1.2 M sorbitol in 0.12 M K2HPO4/0.033 M citric acid, pH 5.9. Fixed cells were digested with 0.1 mg/ml zymolyase-100T (US Biological) and 1/10 volume of glusulase (PerkinElmer) at 30C for 15 min, washed once, and resuspended in 1.2 M sorbitol in 0.12 M K2HPO4/0.033 M citric acid, pH 5.9. Primary antibodies were anti-HA monoclonal antibody (HA.11; 1:500) and anti-tubulin (Abcam; 1:500). Secondary antibodies were: anti-mouse Cy3 (for HA) and anti-rat FITC (for tubulin). Cells were resuspended in DAPI (1 mg/ml). 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.

Preparation: formaldehyde fixed tissue
Relation to intact cell: whole mounted tissue
Item type: recorded image
Imaging mode: widefield illumination; fluorescence microscopy
Parameter imaged: fluorescence emission
Source of contrast: distribution of a specific protein; compartmentalization of stain or label
Visualization methods: HA peptide tag; 4',6-diamidino-2-phenylindole (DAPI); primary antibody plus labeled secondary antibody; Cy3; Fluorescein (FITC)
Processing history: unprocessed raw data
Data qualification: Raw;spatialmeasurements;intensitiesquantitation

• Cell Image Library Group ID: 9034

• Saccharomyces cerevisiae

• Cytoplasm
• Microtubule
• Nucleus
• Spindle pole body
• Tubulin complex

• Cell cycle
• GTP binding
• GTPase activity
• Microtubule-based process
• Mitotic sister chromatid segregation
• Nucleotide binding
• Protein dephosphorylation
• Protein tyrosine/serine/threonine phosphatase activity
• Regulation of exit from mitosis
• Structural constituent of cytoskeleton
• W303

• data
• image

• No linguistic content; Not applicable

Samplenumber: 13865

Doi: https://doi.org/10.6075/J0R78CXC

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL13865

Reference

• J Cell Biol. (2011) 192: 599-614. https://www.ncbi.nlm.nih.gov/pubmed/?term=21321099

Other resource

• BioStudies (previously JCB DataViewer): https://www.ebi.ac.uk/biostudies/studies/

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2022-08-12