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SRX10640875: GSM5253865: 501-KO5-2-MITF; Homo sapiens; ChIP-Seq
1 ILLUMINA (NextSeq 500) run: 46.3M spots, 3.5G bases, 1.3Gb downloads

Submitted by: NCBI (GEO)
Study: PBAF complex deficiency in melanoma mediates BAF occupancy and transcription factor dynamics (Chromatin)
show Abstracthide Abstract
ARID2 is an essential subunit of the SWI/SNF PBAF chromatin remodeler and is highly mutate in melanoma. To elucidate the role of ARID2 in melanoma biology and chromatin structure we utilized CRISPR Cas9 methodology to generate isogenic ARID2 WT and KO melanoma clonal cell lines. We further map the genomic localization of several SWI/SNF subunits, open and repressed chromatin markers, and multiple transcription factors to characterize how loss of the PBAF subcomplex alters chromatin accessibility and the melanoma transcription factor network. Finally, we characterized the transcriptional changes produced by PBAF depletion. Overall design: Chromatin characterization of ARID2 KO cell lines
Sample: 501-KO5-2-MITF
SAMN18806628 • SRS8737104 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: NextSeq 500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: XL ChIP-seq: For ARID2, SS18, BRG1, TEAD4, FOSL2, CTCF, and MITF cells were cross-linked with 2 mM disuccinimidyl glutarate (DSG, Pierce 20593) in PBS for 45 min followed by 1% FA for 10 min at RT. For H3K27ac and H3K27me3 cells were cross-linked with 1% FA for 10 min at RT. ChIP was performed as previously described (Fontanals-Cirera B, Hasson D, Vardabasso C, Di Micco R et al. Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene. Mol Cell 2017 Nov 16;68(4):731-744.e9) ChIP-seq: Libraries for ChIP-seq were done as previously described (Fontanals-Cirera B, Hasson D, Vardabasso C, Di Micco R et al. Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene. Mol Cell 2017 Nov 16;68(4):731-744.e9) ATAC-seq: For all ATAC-seq libraries, 50k cells were harvested, tagmented with 2.5-5uL Nextera Tn5 Transposes from nextera for 30 min, amplified up to 10 cycles and purified essentially as previously described in ATAC protocols. Purified libraries were then size selected on 2% agarose gel (150-700bp).
Experiment attributes:
GEO Accession: GSM5253865
Links:
Runs: 1 run, 46.3M spots, 3.5G bases, 1.3Gb
Run# of Spots# of BasesSizePublished
SRR1427975846,311,7423.5G1.3Gb2022-04-06

ID:
14137746

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