GSM8194270: Baseline, Csf1r∆FIRE/∆FIRE, FIRE_NI_KO R2; Mus musculus; RNA-Seq
Heart tissue was dissected into remote and ischemic tissue as described above and minced into small pieces using forceps and a scalpel. When comparing baseline and I/R injury in FACS analysis, basal heart tissue was used for comparison with remote tissue and apical heart tissue for comparison with ischemia tissue. After enzymatic digestion (Collagenase XI 1200 U/mg, Collagenase I 125 U/mg, Hyaluronidase 500 U/mg, DNase I 1836 U/mg; Sigma Aldrich®) for 30 min at 37°C cells were washed and incubated with purified anti-CD16/32 (FcγRIII/II; dilution 1/50) for 10 min. After sorting, cells were proceeded for single cell capture, barcoding and library preparation using Chromium Next GEM single cell 3' (v3.1, 10x Genomics) according to manufacturer's specifications. Pooled libraries were sequenced on an Illumina HiSeq1500 sequencer (Illumina, San Diego, USA) in paired-end mode with asymmetric read length of 28+91 bp and a single indexing read of 8bp.