Single nuclei were isolated from 4 samples for single-nucleus RNA-seq as previously described (60 (link)), using EZ Lysis buffer workflow with slight modifications. Briefly, tissue samples were thawed in PBS and cut into pieces < 0.5 cm. Approximately 35 mg of tissue were poured in a glass Dounce tissue grinder (Sigma, cat. no. D8938) and homogenized 25 times with pestle A and 25 times with pestle B in 1.5 mL of ice-cold nuclei EZ lysis buffer. Samples were then incubated on ice for 5 min with an additional 3 mL of cold EZ lysis buffer. Nuclei were centrifuged at 500 g for 5 min at 4 °C, washed with 5 mL ice-cold EZ lysis buffer and incubated on ice for 5 min. After centrifugation, the nucleus pellet was washed with 5 mL of Nuclei Wash buffer containing 1× PBS, 0.1%, non-acetylated BSA (Thermo AM2618) and 200 units/mL RNase inhibitor (NEB M0307L). Isolated nuclei were resuspended in 2 mL of Nuclei Suspension Buffer containing 1× PBS, 1% non-acetylated BSA (Thermo AM2618) and 200 units/mL RNase inhibitor (NEB M0307L), filtered through a 70 µm and then a 30 μm MACS SmartStrainers (Miltenyibiotec 130–098-462 & 130–098-458), and counted under microscope using C-chip disposable hemocytometer. A final concentration of 1,000 nuclei per µL was used for loading on a 10x channel.