GSM8045574: wt_1mo_m; Mus musculus; RNA-Seq
Four P30 retinas, one WT male, one WT female, one NLGF male, and one NLGF female WT were extracted according to the published procedure (Fadl et al., 2020). Briefly, the retinas were dissected in fresh and cold 1X HBSS and individually transferred to a 5 ml polypropylene round-bottom tube containing 1 ml of digestion solution. The individual retinas were then incubated at 8 °C for 40 min followed by a second incubation at 28 °C for 10 min. After the incubation steps, the retina should have remained morphologically intact. Mechanical trituration of the retina was performed in 700 µl of prewarmed (10 min at 28 °C) inactivation solution. Trituration was stopped when the retina was visibly dissociated, and 700 µl of ice-cold washing solution was layered under the cell suspension. Cell suspensions were centrifuged using a swing-bucket rotor at 200 ×g for 5 min at 4 °C. After the supernatant was removed, the cells were resuspended in 500 µl of DPBS containing 0.04% BSA and passed through a 40 μm cell strainer. Single cell suspensions from retinal dissociation were counted and loaded onto a droplet based 10x Chromium controller to perform single cell partitioning and barcoding.