title: GSM4005124: F9; Mus musculus; RNA-Seq
library_construction_protocol: Cardiac cells were collected at different time points after myocardial infarction (no infarct/Day 0: n=5; Day 1: n=5; Day 3: n=5, Day 5: n=5; Day 7: n=3) as described above. Cell suspensions were incubated for 10 minutes at 4°C with 10μg/ml Fc Block (purified rat anti-mouse CD16/CD32, Biolegend TruStain FcX anti-mouse) to block non-specific binding of antibodies to Fc Receptors and then incubated at 4°C for 30 minutes with anti-CD45-PE (1:300, Thermofisher Scientific, Clone 30-F11), anti-Ly6G-V450 (1:300, BD Biosciences, Clone 1A8) and Fixable Viability Staining e780 (1:1000, Thermofisher Scientific). All staining and washing steps were performed in PBS containing 1% FCS, except the last wash and sorting that were made in PBS supplemented with 0.04% BSA (Sigma Aldrich, SRE0036). Right before sorting, cells were passed through a 40µm cell strainer. In this experiment, a neutrophil reduction step was performed to increase the proportion of monocyte/macrophages at early time points. Viable CD45.2 (intravenous)-negative/CD45+ cells were sorted using a FACS-Aria III (BD Biosciences) with a 100µm nozzle, counted and loaded into the 10X Genomics Chromium. Cells were loaded in the 10X Genomics Chromium at concentrations recommended by the manufacturer. Libraries were prepared with Chromium Single Cell 3' Reagents Kit v2 Chemistry.
attributes.tag: GEO Accession
attributes.value: GSM4005124