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| Status |
Public on Apr 20, 2022 |
| Title |
P0_ERa_neg_female_E2_CR |
| Sample type |
SRA |
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| Source name |
Limbic system
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| Organism |
Mus musculus |
| Characteristics |
tissue: Brain
Sex: Female
treatment: Estradiol
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| Treatment protocol |
Animals were injected subcutaneously with 5 ug estradiol benzoate or vehicle control (corn oil) 4 hours prior to brain dissection.
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| Growth protocol |
P0 Esr1Cre/+; Sun1-GFP/+ female mice were treated for 4 hr.
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| Extracted molecule |
genomic DNA |
| Extraction protocol |
Tissue (4-5 animals pooled per replicate) was homogenized 15x with a loose pestle in a glass homogenizer containing Homogenization Medium (250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 20 mM Tricine-KOH, 1 mM DTT, 0.15 mM spermine, 0.5 mM spermidine, 1X Roche EDTA-free protease inhibitor cocktail, pH 7.8). 0.3% IGEPAL CA-630 was added, and the tissue was further dounced 5x with a tight pestle. After douncing, the homogenate was filtered through a 40 µm strainer.The nuclei were diluted 2:1 with cold, supplemented homogenization buffer, and 2mM EDTA was added. The sample was immediately sorted using the Sony SH800S Cell Sorter (purity mode) with a 100-μm sorting chip. 150,000 GFP+ nuclei were collected into CUT&RUN wash buffer. GFP- events were collected into CUT&RUN wash buffer, and 150,000 nuclei were subsequently counted on the Countess II FL Automated Cell Counter for ERα- and IgG negative control CUT&RUN.
Takara SMARTer ThruPlex DNA-seq Kit
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| Library strategy |
OTHER |
| Library source |
genomic |
| Library selection |
other |
| Instrument model |
Illumina NextSeq 500 |
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| Description |
150,000 nuclei
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| Data processing |
library strategy: CUT&RUN
Paired-end reads were trimmed to remove low-quality basecalls and adapters (cutadapt -q 30).
Trimmed reads were aligned to mm10 using Bowtie2 with the following flags: --dovetail --very-sensitive-local --no-unal --no-mixed --no-discordant --phred33
Duplicate reads were removed using Picard MarkDuplicates. Read pairs with MAPQ < 40 (samtools view) and fragment length > 120 bp were removed (deeptools alignmentSieve). Read pairs aligning to the mitochondrial genome or incomplete assemblies were also removed (samtools view).
Peaks were called using MACS2 callpeak (q<0.01). Filtered BAM files were normalized by coverage (bamCoverage -bs 1 --normalizeUsing CPM) to generate bigwig tracks.
Genome_build: mm10
Supplementary_files_format_and_content: Bigwig files represent normalized genomic coverage. narrowPeak files represent regions of enriched signal.
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| Submission date |
Jan 12, 2022 |
| Last update date |
Apr 20, 2022 |
| Contact name |
Melody Wu |
| E-mail(s) |
mwu@cshl.edu
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| Organization name |
Cold Spring Harbor Laboratory
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| Street address |
1 Bungtown Rd
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| City |
Cold Spring Harbor |
| ZIP/Postal code |
11724 |
| Country |
USA |
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| Platform ID |
GPL19057 |
| Series (2) |
| GSE144716 |
Gene regulation by gonadal hormone receptors defines brain sex differences-[CUT&RUN] |
| GSE144718 |
Gene regulation by gonadal hormone receptors defines brain sex differences |
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| Relations |
| BioSample |
SAMN24900548 |
| SRA |
SRX13755766 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSM5814052_P0_ERa_female_E2_GFP_neg.bw |
110.5 Mb |
(ftp)(http) |
BW |
| GSM5814052_P0_ERa_female_E2_GFP_neg.narrowPeak.gz |
1.4 Kb |
(ftp)(http) |
NARROWPEAK |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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