GSM9662022: GFP+ and CD45+ cells; Mus musculus; RNA-Seq
For FACS, retinas were dissociated into single cells using the Papain kit from Worthington. Retinas were incubated in a solution of papain and deoxyribonuclease (DNase) for 10 minutes at 37oC, followed by trituration. To stop the reaction, an equal volume of ovomucoid was added. Cells were then spun at 4oC at 300g for 10 min, resuspended in Neurobasal solution and passed through a 35um filter. Using a SONY SH800s cell sorter, FACS was performed on GFP+ cells for reprogramming studies and CD45+ stained-cells for immune response studies. After FACS, the cells were spun down at 300g at 4oC. Cells were loaded on the 10x Genomics single cells chip and processed according to the Chromium Single Cell 3' Reagents Kit V4 user guide. Libraries were sequenced using the Illumina NextSeq 2000 P3 XLEAP-SBS" Reagent Kit. Each library is generated from pooled cells across multiple mice.