Skip to main content
. Author manuscript; available in PMC: 2022 Sep 7.
Published in final edited form as: Cell Metab. 2021 Aug 20;33(9):1853–1868.e11. doi: 10.1016/j.cmet.2021.08.002

Figure 3: Adipocytes undergoing energetic stress release respiration-competent but oxidatively damaged mitochondria in sEVs.

Figure 3:

A, sEVs released from dox-treated, in vitro-differentiated, control and adipo-FtMT adipocytes. B-D, sEV production by in vitro-differentiated wild type adipocytes treated with the indicated compounds. Media was harvested for sEV quantification after 24 hours of treatment (B and D) or the timepoints indicated in C. E, Western blot was performed for mitochondrial proteins in sEVs isolated from the serum of control or adipo-FtMT mice on dox-HFD for 3 weeks (representative of n=3). F, Optiprep density gradient purification of serum sEVs isolated from adipo-FtMT mice fed dox-HFD for 3 weeks. G and H, Mitochondrial proteins were assessed in sEVs from wild type in vitro-differentiated adipocytes (G; representative of n=3) and dox-treated in vitro-differentiated control and adipo-FtMT adipocytes (H; representative of n=5). I and J, Western blot and densitometry for mitochondrial proteins in sEVs produced by primary in vitro-differentiated adipocytes treated as shown. K, Mitochondrial protein content in serum sEVs from lean and obese (20 weeks HFD) wild-type mice. L and M, Seahorse Flux Analysis of isolated serum sEVs from control or adipo-FtMT mice fed dox-HFD for 3 weeks. N, Protein carbonylation (PC) assay on isolated sEVs from control and adipo-FtMT mice on dox-HFD for 3 weeks or conditioned media from adipocytes treated as indicated. sEVs recovered from equal amounts of media or serum were loaded into the SDS-PAGE gels. Abbreviations: oligo, oligomycin; Anti A, antimycin A; PA, palmitate; Ad, adipocyte. Data are presented as mean ± s.e.m. *P < 0.05, ** P < 0.01, *** P < 0.001. See also Figure S5 and S6.