Adipose tissues were collected and washed in PBS before subjected to lipolysis assays. For isoproterenol stimulation, adipose tissues were cut into small tissue pieces and incubated in serum‐free DMEM–High Glucose (Sigma, D5796) with 2% fatty acid‐free BSA (Sigma, 126579) in the absence or presence of 10 μM isoproterenol (Sigma, I6504) for the indicated time. TNFα‐induced lipolysis was induced as previously described (Ju et al, 2019 (link)). In brief, small adipose tissue pieces were cultured in DMEM–High Glucose for 24 h in the absence or presence of 100 ng/ml recombinant TNFα (Peprotech, 315‐01A) and then transferred into serum‐free DMEM containing 2% fatty acid‐free BSA for 3 h. Supernatants were collected and FFA concentration normalized to adipose tissue input.
Richter F.C., Friedrich M., Kampschulte N., Piletic K., Alsaleh G., Zummach R., Hecker J., Pohin M., Ilott N., Guschina I., Wideman S.K., Johnson E., Borsa M., Hahn P., Morriseau C., Hammock B.D., Schipper H.S., Edwards C.M., Zechner R., Siegmund B., Weidinger C., Schebb N.H., Powrie F, & Simon A.K. (2023). Adipocyte autophagy limits gut inflammation by controlling oxylipin and IL‐10. The EMBO Journal, 42(6), e112202.
Other organizations :
University of Oxford, John Radcliffe Hospital, University of Wuppertal, Max Delbrück Center, Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Humboldt-Universität zu Berlin, Freie Universität Berlin, Cardiff University, MRC Human Immunology Unit, MRC Weatherall Institute of Molecular Medicine, University of California, Davis, University Medical Center Utrecht, Nuffield Orthopaedic Centre, University of Graz
Positive control: Adipose tissue pieces incubated with isoproterenol (10 μM) or TNFα (100 ng/ml)
Negative control: Adipose tissue pieces incubated without isoproterenol or TNFα
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