GSM9589025: retina, snRNA-seq, Olig2-cre;Piezo2f/f cKO, P120, Sample 13; Mus musculus; RNA-Seq
After nuclei isolation using Dounce homogenizers and ultracentrifugation at 500g for 5 min,  pelleted nuclei were resuspended in staining buffer containing primary antibodies targeting NeuN and RBPMS to detect RGCs for 15 minutes at 4 °C. Nuclei were washed, centrifuged at 500 × g for 5 minutes, stained with secondary antibodies and sorted using a BD FACSDiva device. Sorted NeuN+/RBPMS+ nuclei were pelleted at 500 × g for 5 minutes at 4 °C, resuspended, counted in a Neubauer chamber and adjusted to a concentration of ~600 nuclei µl–1. Nuclei were then loaded onto a microfluidic platform (Chromium X Series controller) aiming for 8,000 nuclei per reaction. Libararies were generated according to the 10x Next GEM 3' Kit v.3.1 protocol.