Journal: Cell reports

Article Title: Extracellular vesicles are carriers of adiponectin with insulin-sensitizing and anti-inflammatory properties.

doi: 10.1016/j.celrep.2023.112866

Figure Lengend Snippet: Figure 1. Adiponectin is the most enriched adipokine in VAT-derived EVs (A and B) Lean (Ctrl) and obese (Ob) VAT-derived lEV and sEV mode size comparison. Size-distribution curves (A) and mean mode size (B). Ctrl and Ob EV subtype size-distribution curves (A) are represented by plain or dashed lines, respectively, and are presented as the mean ± SEM (n = 4–6 independent EV preparations for each condition). (C and D) EV marker analysis from lean (Ctrl) or ob/ob (Ob) VAT explant-derived EV subpopulations. A significant enrichment in CD63 was observed for obese VAT-derived sEVs (D). (E) lEV and sEV secretion from VAT, SAT, and BAT explants. EV number secreted per gram of AT per 24 h is presented. (F) Increased lEV and sEV secretion from mouse VAT with obesity. EV secreted by total mouse VAT is presented. (G) EV secretion of human omental, mesenteric (Mesent.), and subcutaneous (Subcut.) AT collected from obese subjects. Secretion of EVs is presented as the number of EVs secreted per gram of human AT.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Exosome isolation kit Pan, mouse Miltenyi Biotec Cat #120-041-065 Fetal Calf serum (FCS) Eurobio Cat #CVFSVF00-01 Insulin 100UI/mL Lilly Cat #HI0210 Zeocin Thermofisher Scientific Cat #R25001 Palmitate Sigma Cat #P-0500 Proteinase K Sigma Aldrich Cat #P2308 PMSF Santa Cruz Cat #SC24948 ECBM PromoCell Cat #C22210 Fetal Bovine serum Gibco BRL Cat #10270-106, Lot 42G2078K Fluoromount-G Invitrogen Cat #00-4958-02 Glutaraldehyde 25% solution Electron microscopy Sciences Cat #16220 IRDye 800CW NHS ester LICOR Cat #92970021 Mini-Protean TGX gels Bio-Rad Cat #4568083, #4568086, Mowiol Calbiochem Cat #475904 OptiprepTM density gradient medium Sigma Aldrich Cat #D1556 Odyssey blocking buffer (TBS) LI-COR Biosciences Cat #927-50010 Paraformaldehyde 32% Electron microscopy Sciences Cat #15714 Running buffer Interchim Cat #91495E 4X Laemmli sample buffer Bio-Rad Cat #161-0747 Triton X-100 Sigma Cat #T8787 Trans-blot Turbo transfer reagents Bio-Rad Cat #10026938, #1704158, #1704271 Trypsin 0.05% Corning Cat #25-051-CI RNAiMax Lipofectamine Invitrogen Cat #12323563 High-fat diet (HFD) 61% of the energy from fat (D12492 Premix AIN) Safe diet Cat# SAFE 233 HF Critical commercial assays DC-protein assay BioRad Cat #5000111 Mouse adipokine array kit R&D systems Cat #ARY013 Mouse and human adiponectin/ Acrp30 DuoSet ELISA R&D systems Cat #DY1119 and DY1065 Human magnetic Luminex set for adiponectin Bio-Rad Cat #171A7002M Ultrasensitive mouse Elisa kit Crystal Chem Cat #90080 Element RC liver rotor Scil animal care company Cat #AW00199 Glucometer strips for glucometer AccuCheck Guide Roche Cat #08721106001 Clinicells 25cm3 GEDTECH Cat #00108 Experimental models: Cell lines HEK 293 ATCC Cat #CRL-1573 HepG2 cells ATCC Cat #HB-8065 Phoenix-AMPHO cells ATCC Cat #CRL-3213 Experimental models: Organisms/strains Ob/Ob mice Charles River B6.Cg-Lepob/J, stock N 000632, JAXTM mice strain RCL-ZsGreen The Jackson Laboratory B6.Cg-Gt(ROSA)26Sortm6(CAG ZsGreen1)Hze/J, stock N 007906, JAXTM mice strain (Continued on next page) Cell Reports 42, 112866, August 29, 2023 17

Techniques: Derivative Assay, Comparison, Marker