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SRX22550716: GSM7904806: BALBc_Ctrl_2; Mus musculus; ATAC-seq
1 ILLUMINA (NextSeq 2000) run: 110.1M spots, 11.2G bases, 3.4Gb downloads

External Id: GSM7904806_r1
Submitted by: National Institutes of Health
Study: Genetic variation in IL-4 activated tissue resident macrophages alters the epigenetic state to determine strain specific synergistic responses to LPS [ATAC-seq]
show Abstracthide Abstract
IL-4 activates tissue resident macrophages (TRMs) to mediate tissue repair and clearance of nematode infections in mice, but most studies have been performed on the C57BL/6 background. The natural genetic variation between C57BL/6 and BALB/c mouse strains can result in differences in allergic responses, parasite resistance, monocyte to macrophage conversion and response to IL-4 activation. Here, we investigated in vivo IL-4 activation in TRMs of the peritoneal cavity from C57BL/6 and BALB/c mice and find that C57BL/6 TRMs are surprisingly more transcriptionally responsive to IL-4 stimulation, with greater association of induced genes with super enhancers, induced enhancers and topologically associating domains (TAD) boundaries. IL-4-directed epigenomic remodeling showed NF-?B motif enrichment only in C57BL/6 TRMs. This resulted in an enhanced synergistic response upon in vitro lipopolysaccharide (LPS) exposure in IL-4 activated C57BL/6 TRMs but not for BALB/c, despite unstimulated BALB/c TRMs having a stronger transcriptional response to LPS than C57BL/6 TRMs. Single cell RNA sequencing (scRNAseq) analysis of mixed bone marrow chimeric mice indicates that transcriptional differences between C57BL/6 and BALB/c TRMs is cell intrinsic within the same tissue environment. Hence, genetic variation alters IL-4-induced cell intrinsic epigenetic reprogramming between C57BL/6 and BALB/c TRMs to regulate the magnitude of synergistic responses to LPS exposure. Overall design: 8 samples were anlysied (Ctrl and IL4 treatment in each strain, 2 replicates per samples) UPDATE: [09-30-2024] The GSM7904801_BL6_Ctrl_1.genrich.RPM.bw file was renamed to GSM7904804_BL6_IL4_2.genrich.RPM.bw, and vice versa.
Sample: BALBc_Ctrl_2
SAMN38286061 • SRS19559932 • All experiments • All runs
Organism: Mus musculus
Library:
Name: GSM7904806
Instrument: NextSeq 2000
Strategy: ATAC-seq
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: 100,000 macrophage cells were subjected to lysis using 100 µl of Lysis buffer (containing 10 mM Tris-Cl at pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, and 0.1% Tween-20). Following a centrifugation at 500g for 10 minutes at 4°C, the resulting nuclei were reconstituted with 50 µl of a transposition mixture (FC-121-1030, Illumina; comprising 1X Tagment DNA buffer, Tn5 Transposase, and nuclease-free H2O), and this mixture was then incubated for 30 minutes at 37°C in a thermomixer. The transposed DNA was subsequently purified using MinElute columns (28004, QIAGEN) and then amplified using Nextera sequencing primers and the NEB high-fidelity 2X PCR master mix for 11 cycles (M0541, New England Biolabs). PCR-amplified DNA was purified using MinElute columns ATAC-seq samples were pooled and sequenced on NextSeq 2000 P2 using PI lab prepared library and paired-end sequencing.
Runs: 1 run, 110.1M spots, 11.2G bases, 3.4Gb
Run# of Spots# of BasesSizePublished
SRR26855508110,127,07711.2G3.4Gb2023-12-29

ID:
30546183

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