title: GSM8128514: 24W_scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: P5, 12w, and 24w Nestin-EGFP mice were euthanized, and each brain was promptly harvested. The DG was quickly microdissected under a dissection scope and minced with a scalpel. The dissociated DG was transferred into prewarmed papain (25 U/mL) solution and incubated at 33 ℃ for 45 min. The suspension was then washed with 2 mL Minimum Essential Medium α (Gibco) containing 5% BSA to stop enzyme activity, mechanically dissociated by gentle pipetting with a fire-polished Pasteur pipette, and centrifuged at 130 × g for 5 min. The cell pellet was then resuspended in 2 mL of Hanks' balanced salt solution (HBSS) containing 250 U/mL DNase and centrifuged at 130 × g for 5 min, after which the cells were suspended in HBSS. The cells were sorted on a FACS Aria II (BD Biosciences), with 7-AAD (559925, BD Pharmingen) added to exclude nonviable cells from the analysis. Cells dissociated from the DG of WT mice were used to draw gates for EGFP+ and EGFP− cell populations. The EGFP+ cells were sorted into N2 medium and centrifuged at 200 × g at 4 ℃ for 10 min. The supernatant was carefully removed, leaving 30 µL to resuspend the cell pellet. One microliter of cells was mixed with 10 µL Trypan Blue to determine cell viability. For scRNA-seq and scATAC-seq analyses, 700–1200 cells/µL were used. Libraries were prepared according to the manufacturer's instructions (10X Genomics).