421-Takenouchi-AgedYoungOocyte/Fig5G_Rec8Antibody_20230118-1-L16
Author reviewed
Time-lapse and z-series images of H2B-SNAP and tdTomato-CENPC in REC8 TRIM-Away oocytes treated with anti-REC8 antibody
Dataset ID
Fig5G_Rec8Antibody_20230118-1-L16
Title
Time-lapse and z-series images of H2B-SNAP and tdTomato-CENPC in REC8 TRIM-Away oocytes treated with anti-REC8 antibody
Description
Time-lapse and z-series images of H2B-SNAP and tdTomato-CENPC in REC8 TRIM-Away oocytes treated with anti-REC8 antibody. By TRIM-Away method, Rec8 was knocked down by the antibody, and cohesin was partially depleted from the oocyte.
Folder Name
Fig5G_Rec8Antibody_20230118-1-L16
Dimensions
256x256x17x3x200
Biosample
Description
From prometaphase to anaphase oocyte from 7 to 9 week-old-mouse, in which chromosomes and kinetochores were visualized. For Rec8 TRIM-Away experiment, anti-REC8 antibody was injected.
Extrinsic Variable
To visualize chromosomes and kinetochores, in vitro
transcribed mRNAs for H2B-SNAP and
tdTomato-CENPC (centromere
protein C) were injected. For TRIM-Away method, 0.38 pg TRIM21 and 0.19 to 0.23 pg anti-REC8 antibody were injected.
Ontology
Biological Process
female meiosis chromosome segregation, (
GO:0016321)
Genetics
Protein / Probe
Histone H2B type 1-J
Method / Genotype
injection
Protein / Probe
Centromere protein C
Method / Genotype
injection
Treatments
No treatment records.
Imaging Method
Detection Method
photomultiplier tube (PMT)
Visualization Method
H2B with SNAP-tag was visuallised with SNAP-Cell 648-SiR. To visualize kinetochores, centromere
protein C was tagged with tdTomato.
Illumination Method
single point scanning
Source of Contrast
distribution of a specific protein
Imaged Parameter
fluorescence emission
Sample Preparation Method
living tissue
Instrument
Objective
Zeiss 40×C-Apochromat 1.2NA