Valerio-Santiago, Mauricio; Monje-Casas, Fernando (2021). CIL:13875, Saccharomyces cerevisiae. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0FN14X5

3HA-Bfa1 (red) localizes to both spindle pole bodies (SPBs) in anaphase in dyn1Δ cells with constitutive targeting of Tem1 to both SPBs. However, the constitutive presence of Tem1 on the SPBs impaired the spindle position checkpoint in the dyn1Δ cells. Anaphase was determined by spindle morphology (tubulin, green) and nuclear morphology (DAPI, blue). A DIC image is also shown (hidden). Image is Fig S3 in J Cell Biol. (2011) 192: 599-614.

• 2021

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

Cells (MATa tem1::KanMX ura3::eGFP-CNM67–TEM1::URA3 dyn1::URA3 bfa1::3HA-BFA1) grown in rich media at 14C for 24 hours 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; differential interference contrast microscopy
Parameter imaged: fluorescence emission; optical path length gradient
Source of contrast: distribution of a specific protein; compartmentalization of stain or label; boundaries between regions with different refractive index
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
• Cytoskeleton
• Dynein complex
• Microtubule
• Nucleus
• Spindle pole body
• Tubulin complex

• ATP binding
• Cell cycle
• Cell division
• Establishment of mitotic spindle orientation
• GTP binding
• GTPase activator activity
• GTPase activity
• Microtubule motor activity
• Microtubule-based movement
• Microtubule-based process
• Mitosis
• Mitotic cell cycle spindle orientation checkpoint
• Mitotic sister chromatid segregation
• Mitotic spindle elongation
• Nuclear migration along microtubule
• Nucleotide binding
• Protein binding
• Regulation of exit from mitosis
• Small GTPase mediated signal transduction
• Structural constituent of cytoskeleton
• W303

• data
• image

• No linguistic content; Not applicable

Samplenumber: 13875

Doi: https://doi.org/10.6075/J0FN14X5

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL13875

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