Nuclei were isolated from 150-250 mg of tissue, which was minced in 1 mL of nuclear extraction buffer (130-128-024, Miltenyi) and transferred into GentleMACS tubes. The h_tumor_01 protocol was run on the GentleMACS three times. Then, tubes were placed on ice for 10 minutes. The homogenate was then passed through a 70 mM filter and washed with 3 mL of nuclei extraction buffer. Samples were centrifuged at 500 x g for 10 minutes at 4°C. Supernatant was removed, and nuclear pellet was resuspended in 2 mL 1% BSA in PBS. Samples were then filtered using a 30 mM filter and rinsed with 2 mL 1% BSA in PBS Samples were centrifuged at 500 x g for 10 minutes at 4°C. Pellets were resuspended in 500 mL 1% BSA in PBS and filtered once more through a pre-wet 20 mM filter. Filters were washed with 500 mL 1% BSA in PBS. Samples were centrifuged at 500 x g for 10 minutes at 4°C, supernatant removed, and nuclear pellet resuspended in 50 mL 1% BSA in PBS. Nuclei were then counted at the University of Michigan Advanced Genomics Core using a Logos cell counter. Probes were hybridized, samples were pooled and processed according to manufacturer instructions for 10X Genomics Chromium Fixed RNA Profiling Reagent Kits for Multiplexed Samples (PN 1000568). Final library quality was assessed using the LabChip GXII HT (PerkinElmer) and libraries were quantified by Qubit (ThermoFisher). Libraries were generated according to the manufacturer's instructions (10x 3' v3 for MacDougald_Adipose_2025.1, MacDougald_Adipose_2025.2, and MacDougald_Adipose_2025.3 and 10x 3' v4 for MacDougald_Adipose_2025.4, MacDougald_Adipose_2025.5, MacDougald_Adipose_2025.6).