GSM5350881: P10 WT rep C7; Mus musculus; RNA-Seq
Four retinas from P6 or P10 and WT or Alk5iEKO pups were dissected and dissociated with a modified version of the Neural Tissue Dissociation kit (P) (Miltenyi Biotec, Cat#130- 092-628). The retinas were digested in buffer P supplemented with DNase I (2000 U/mL, Qiagen #79254) for 5 min. The digestion was stopped by adding FBS (10% final) and the cells were immediately filtered through a 70 μm cell strainer. After centrifugation (300g for 3 min), the cells were resuspended in PBS supplemented with 2mM EDTA and 0.5% BSA. The single cell suspension was enriched for ECs using CD31 MicroBeads (Miltenyi Biotec, Cat#130-097-418) according to the manufacturer's instructions. Single cell suspensions of retinal cells were resuspended in PBS containing 0.04% ultra556 pure BSA. Libraries were prepared using the Chromium Single Cell 30 Reagent Kits v3.1 (10x Genomics; Pleasanton, CA, USA) according to the manufacturer's instructions. scRNA-seq