title: GSM5466303: Sham_m_S1_5; Mus musculus; RNA-Seq
library_construction_protocol: Each block of tissue was homogenised using a dounce glass tissue homogeniser. Mononuclear cells were separated through Percoll (GE Healthcare Life Sciences) 37%/70% gradient centrifugation. Mononuclear cells were isolated from the interface and stained on ice for 30 min with combinations of BV650 rat anti-mouse CD11b (marking myeloid cells; 1:300), FITC rat anti-mouse Ly6C (marking monocytes/macrophages; 1:300) and APC rat anti-mouse 4D4 (marking resident microglia; 1:1000) in blocking buffer containing 0.2% bovine serum albumin (BSA, Sigma-Aldrich) in HBSS. Cell sorting was performed using FACSAriaIII cell sorter (Becton Dickson). Microglial cells were identified as CD11b+ Ly6C- 4D4+ cDNA libraries were generated from sorted cells using the Smart-seq2 protocol