title: GSM4718812: E12_G4_TNT_rep1; Mus musculus; ChIP-Seq
library_construction_protocol: GATA4 bioChIP-seq:  For each bioChIP-seq of GATA4, 15-20 E12.5 heart ventricles were harvested and crosslinked in 1% formaldehyde in PBS for 10 minutes at room temperature. Chromatin was fragmented using a microtip sonicator (Qsonica, S-3000) at 65% amplitude for 8 minutes. GATAfb and bound chromatin was pulled down by incubation with streptavidin beads (ThermoFisher Scientific, Cat#11206D) GATA4 bioChIP-seq:   Libraries were constructed using the KAPA HyperPrep ChIP-seq library preparation kit (Roche, cat# 07962347001). Sequencing (75 nt single end) was performed on an Illumina NextSeq 500. EC ATAC-seq:  For ATAC-seq of Tie2-Cre cell lineages, E12.5 Rosa26fsTrap;Tie2Cre mouse heart ventricles were dissected in ice cold PBS and dissociated to single cells with Neonatal Cardiomyocyte Isolation kit (Cellutron, cat#nc-6031). 40K GFP+ cells were sorted (BD FACSMelody) for each ATAC-seq library sample. For ATAC-seq of Cdh5(Pac)-CreERT2 labeled, Gata4 mutant and control cell lineages, 1mg tamoxifen dissolved in sunflower seed oil was administered to pregnant mice by gavage to induce the activity of Cre at E9.5. E12.5 heart ventricles were dissected in ice cold PBS and dissociated to single cells with the Neonatal Cardiomyocyte Isolation kit (Miltenyi). 20K GFP+ cells were sorted (BD FACSMelody) for each ATAC-seq library sample. EC ATAC-seq:  The optimized-ATACSeq (omni-ATACSeq) libraries were generated from purified cells using the author's recommended lysis steps and transposition protocol EC ATAC-seq:  optimized-ATACSeq (omni-ATACSeq) EC RNA-seq:  E12.5 Rosa26fsTrap;Tie2Cre mouse heart ventricles were dissected in ice cold PBS and dissociated to single cells with Neonatal Cardiomyocyte Isolation kit (Cellutron, cat#nc-6031). 60K GFP+ cells were sorted (BD FACSMelody) for each RNA sample. EC RNA-seq:  RNA was prepared with RNeasy Micro Kit (Qiagen, Cat#74004). The polyadenylated RNA was purified by binding to oligo (dT) magnetic beads (ThermoFisher Scientific, 61005). EC RNA-seq:  RNA-seq libraries were prepared with ScriptSeq v2 kit (Illumina, SSV 21106) according to the manufacturer's instructions.
attributes.tag: GEO Accession
attributes.value: GSM4718812