Quantifying TRL Margination in Vasculature

The margination of TRLs in blood vessels was measured as described previously (31 (link)). Briefly, TRLs were isolated from the plasma of Gpihbp1^–/– mice by ultracentrifugation (d < 1.006 g/mL). Mice were injected intravenously with IRDye680-TRLs and IRDye800-2H8 in saline containing 0.25 mM tetrahydrolipstatin. After 60 s, mice were perfused with 20 mL PBS through the left ventricle, followed by 10 mL 3% PFA in PBS. Tissues were embedded in OCT; 10-μm-thick frozen sections were prepared (6 sections/tissue/mouse) and scanned using an infrared scanner. The TRL signal was normalized to the 2H8 signal.

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Song W., Yang Y., Heizer P., Tu Y., Weston T.A., Kim J.R., Munguia P., Jung H., Fong J.L., Tran C., Ploug M., Beigneux A.P., Young S.G, & Fong L.G. (2023). Intracapillary LPL levels in brown adipose tissue, visualized with an antibody-based approach, are regulated by ANGPTL4 at thermoneutral temperatures. Proceedings of the National Academy of Sciences of the United States of America, 120(8), e2219833120.

Publication 2023

Blood vessels Frozen sections Irdye800 Left ventricle Mouse Plasma Saline Tetrahydrolipstatin Tissue Ultracentrifugation

Corresponding Organization :

Other organizations : University of California, Los Angeles, Rigshospitalet, University of Copenhagen

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Variable analysis

independent variables

Intravenous injection of IRDye680-TRLs and IRDye800-2H8 in saline containing 0.25 mM tetrahydrolipstatin

dependent variables

Margination of TRLs in blood vessels

control variables

Perfusion with 20 mL PBS through the left ventricle, followed by 10 mL 3% PFA in PBS
Tissue embedding in OCT and preparation of 10-μm-thick frozen sections
Scanning of tissue sections using an infrared scanner
Normalization of TRL signal to the 2H8 signal

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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