title: GSM8740252: 10xMulti_Pomc_4_HTO; Mus musculus; OTHER
library_construction_protocol: Tissue preparation, dissociation, and nuclei extraction were described in a STAR Protocol (https://doi.org/10.1016/j.xpro.2023.102595). In this study, the ARH regions were dissected from four-week-old for Pomc-Cre; Sun1/sfGFP and Pomc-Cre; Otpfl/fl; Sun1/sfGFP mice, under a fluorescent dissecting microscope and stored at -80°C. Different genotype samples were separately homogenized with 1 ml homogenization buffer using a Dounce grinder pestle. The homogenate solutions were centrifuged at 500 g for 5 min in a swing bucket rotor, and the supernatants were removed. The nuclei were resuspended with 100 μl of wash buffer and incubated with different TotalSeq Hashtag antibodies against the nuclear pore complex (0.5 μl, MAb414; BioLegend) for 10 min. The nuclei suspensions were washed thrice with 1 ml wash buffer and spun down at 500 g for 5 min. After hashtagging each sample with the antibody, all samples were pooled together, and GFP-positive nuclei were sorted during FACS at the Flow Cytometry Core of UTSW Medical Center. After sorting, the nuclei solution was adjusted to the desired concentration at 500–2,000 nuclei/μl, then proceeded with the 10× Multiome kit at the UTSW Next Generation Sequencing Core.
identifiers.namespace: GEO
identifiers.id: GSM8740252_r1