GSM6222357: LSK, CLP, scRNAseq; Mus musculus; RNA-Seq
Bone marrow cells were harvested from femurs and tibias collected from 3 mice at 12 weeks after sham or CLP surgery, and were resuspended in FACS buffer (PBS supplemented with 2 mM EDTA and 0.5% BSA). Lineage marker-negative cells were first enriched from bone marrow cells using EasySep Mouse Hematopoietic Progenitor Cell Isolation Kit (StemCell Technologies), and then stained with antibodies specific for lineage markers [B220 (clone RA3-6B2), CD19 (clone 1D3), TCR (clone H57-597), CD3 (clone 17A2), CD4 (clone GK1.5), CD8 (clone 53-6.7), CD11c (clone N418), CD11b (clone M1/70), Gr1 (clone RB6-8C5), Ly6C (clone AL-21), F4/80 (clone BM8), TER119 (clone TER-119), NK1.1 (clone PK136)], c-Kit (clone 2B8) and Sca1 (clone D7). FACS sorting was performed on a FACSAria II instrument (BD Biosciences). LSK cells were washed and resuspended in PBS supplemented with 0.04% w/v BSA. Single cell partitioning and cDNA library construction were done using Chromium Single Cell 3' Kit v3 per manufacturer's instruction (10X Genomics).