GSM8581623: ROBO_CD31; Mus musculus; RNA-Seq
Briefly, 8 retinas per group were dissected and dissociated with the Neural Tissue Dissociation kit (P) (Miltenyi Biotec, Cat#130- 092-628). The digestion was stopped by adding BSA (5% final) and the cells were immediately filtered through a 70 mm cell strainer. After centrifugation (600g for 5 min), cells were resuspended in PBS supplemented with 2mM EDTA and 0.5% BSA. The single cell suspension was enriched for ECs and immune cells using CD31 and CD45 MicroBeads (Miltenyi Biotec) according to the manufacturer's instructions. Single cell suspensions of retinal cells were resuspended in PBS containing 0.04% ultra-pure BSA. scRNA-seq  libraries were prepared using the Chromium Single Cell 30 Reagent Kits v3.1 (10x Genomics; Pleasanton, CA, USA) according to the manufacturer's instructions. The target cell recovery for each library was 10,000. Generated libraries were sequenced on an Illumina HiSeq4000.