GSM4670657: blood_and_bone_marrow gene expression S1_L001; Mus musculus; RNA-Seq
Lung and liver were mechanically disrupted and passed through a 70 µm strainer to generate single-cell suspensions followed by percoll gradient centrifugation to enrich CD45+ immune cells. Bone marrows were flushed with buffer (DPBS with 1% BSA) and red blood cells were lysed at room temperature for 5 min. Blood was collected in EDTA tubes and immediately lysed at room temperature and quenched with buffer (DPBS with 1% BSA). The number of cells required from each tissue to give relatively equal numbers of Ly6G+ cells were determined a priori. Blood, bone marrow, liver and lung were labeled with hashing antibodies (Biolegend), pulled and stained with DAPI and Ly6G antibodies (Biolegend) for FACS sorting (BD Influx). 50,000 Ly6G+ cells were sorted with expected 15-30% contribution from each tissue. After FACS sorting, cells were further labeled with CXCR4 and CD62L CITE-seq antibodies (Biolegend) for 20 min on ice. After washing with buffer, cells were counted and immediately processed for single-cell capture and library construction using 10× Genomics Chromium system and 3′ library and gel bead kit v2 (10× Genomics) following standard vendor-guided protocol.
GEO Accession
GSM4670657