ChIP-Atlas: SRX682465

Antigen

Cell type Class: Blood
Cell type: Th2 Cells
MeSH Description: A subset of helper-inducer T-lymphocytes which synthesize and secrete the INTERLEUKINS IL-4; IL-5; IL-6; and IL-10. These cytokines influence B-cell development and antibody production as well as augmenting humoral responses.

source_name: primary CD4+ T cells from spleen and lymph nodes
cell type: in vitro polarized T helper17 cells for 3 days
culture condition: FACS sorted naive CD4+T cells cultured in vitro for 3 days under Th17 condition
strain: C57BL/6J (wild type)
chip antibody: chip antibody: anti-p300, antibody vendor/catalog#: sc-585, Santa Cruz Biotechnology

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: RNA-seq: Total RNA wasprepared from approximately 1 million cells by using mirVana miRNA Isolation Kit(AM1560, ABI). 200 ng of total RNA was subsequently used to prepare RNA-seq library byusing TruSeq SRRNA sample prep kit (FC-122-1001, Illumina) by following manufacturer’s6protocol.The resulted libraries are sequenced using the Illumina HiSeq 2000. Only unique reads of 50 bp were mapped to the mouse genome (mm9) using Tophat and Cufflinks. ChIP-seq: Cells were chemically cross linked by 1% formaldehyde. Cell lysates were made by appropriate sonication and protein-DNA complexes were isolated with antibody. Libraries were prepared according to Illumina's instructions. Briefly, DNA was end-repaired and the blunt, phosphorylated ends were treated with Taq polymerase and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation, DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow celland sequenced on HiSeq 2000.