Exoquick tc precipitation reagent

Isolation of exosomes from adipose tissues was performed following the methods described by Wei et al. (17 (link)) and Yu et al. (18 (link)). Epididymal white adipose tissue (eWAT), visceral white adipose tissue (vWAT), and subcutaneous white adipose tissue (sWAT) from mice were excised and cut into small pieces (5 mm × 5 mm). The shredded tissue was cultured in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% exofree-fetal bovine serum (FBS) (2 mL media/g adipose tissue), and the culture supernatant was collected after 48 h. The supernatant was then centrifuged at 500 × g for 5 min, then 2,500 × g for 15 min, at a temperature of 4°C to remove large debris and dead cells. Exosomes were separated from the resulting supernatant using Exoquick-TC™ precipitation reagent (System Biosciences, Mountain View, CA, United States), per the manufacturer’s instructions. The resulting exosome pellet was resuspended in PBS for application purposes. The identification and quantification of the exosomes were performed using a NanoSight LM10 system (Malvern Panalytical Ltd., Malvern, UK), while the exosomal protein content was evaluated using the Bradford protein assay.

Exosomes were isolated using ExoQuick-TC precipitation reagent (System BioSciences, Palo Alto, CA, USA), as previously described^{17 (link)}.