SA2
The improved nuclear separation method was utilized to isolate nuclei from frozen human white adipose tissue and C-SVF. Sequencing was conducted by SequMed Biotech, Inc. The single-cell nuclear suspension was assessed with a Countess II instrument prior to loading onto the machine, with an anticipated capture of 10,000 nuclei. Single-cell 3' v2 chemistry was employed to produce single-cell barcoded droplets (GEMs), and upon capturing the nuclei, the GEMs  solution was subsequently withdrawn and transferred to PCR tubes for reverse transcription and library construction for sequencing. The "gene expression library" was quantified via a Qubit instrument, and the fragment size of the "gene expression library" was analyzed via Qpcr. Subsequently, Illumina NovaSeq was utilized with a PE150 sequencing strategy for paired-end sequencing, with both reads being 150 bp in length.