title: GSM8679863: shtreated0; Mus musculus; RNA-Seq
library_construction_protocol: A single-cell suspension was prepared from the indicated tissues. For labeling, 40 μl of CD45 microbeads (Miltenyi) were added. After an incubation step (15 min at 4˚C) the antigen-antibody reaction was stopped by adding 800 μl ice-cold PBS. Then, the cell suspension mixture is loaded onto a MACS Column, which is placed in the magnetic field of a MACS Separator. The magnetically labeled CD45+ cells are retained within the column. The unlabeled cells run through; this cell fraction is thus depleted of CD45+ cells, the CD45-negative cells were in the eluent buffer. After removing the column from the magnetic field, the magnetically retained CD45+ cells can be eluted as the positively selected cell fraction and can be used for subsequent experiments. We used GEXSCOPETM Single Cell RNA Library Kit Tissue (Singleron Biotechnologies) to barcode single cells, capture mRNA from isolated single cells, and generate cDNA libraries for scRNA-seq. Then, individual libraries were diluted to 4 ng/μl, and pooled for sequencing on a Novaseq6000 platform (Illumina, San Diego, USA) with 150 bp paired-end reads.