GSM6632267: IL6_KO_Ctrl_S3; Mus musculus; RNA-Seq
IL-6 Wild type (WT, +/+) and IL-6 Knock-out (KO, -/-) mice that were either uninjured (control) or at day 6 post-CVI were anesthetized and perfused using ice cold PBS. Control and CVI-damaged brain tissue was collected using a 4mm biopsy punch (Fisher Scientific; cat#12-460-410). Single cells were dissociated from the punch biopsied brain tissue using a multi tissue dissociation kit 1 (Miltenyi Biotec; cat#130-110-201). After myelin removal using myelin removal beads (Miltenyi Biotec; cat#130-096-733), CD11b+ myeloid cells were positively selected using CD11b microbeads (Miltenyi Biotec; cat#130-093-636). Cell viability and counting were assessed using Acridine orange. Single cell samples in PBS containing 0.05% BSA with viabilities of >80% were used for sequencing. cDNA libraries were prepared from single-cell suspensions following the 10x Genomics 3′ v3 protocol (www.10xgenomics.com) and sequenced on NovaSeq (www.illumina.com) with aim to generate 150M reads/sample for 4,000 cells/sample.