Journal
STAR PROTOCOLS
Volume 3, Issue 4, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.xpro.2022.101665
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Funding
- NIH [R01 HL125838, R01 DK121560, P30 DK056341, T32 HL134635]
- American Heart Association [897628]
- Veterans' Administration [I01 BX003415]
- Longer Life Foundation
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This study provides a protocol for evaluating diet-induced mTORC1 signaling in plaque macrophages in mice, which can be used to study the atherogenic potential of various mTORC1 modulators.
Previous studies have demonstrated that a high-protein diet leads to increased atherosclerosis in mice, and that this adverse effect is caused by activation of macrophage mTORC1 signaling. Here, we provide a detailed protocol for the evaluation of diet-induced mTORC1 signaling in plaque macrophages in athero-sclerosis-prone apolipoprotein E (ApoE) knockout (KO) mice. This protocol includes flow cytometry and immunofluorescence analysis of atherosclerotic macrophages that can be used to study the atherogenic potential of a variety of mTORC1 modulators.For complete details on the use and execution of this protocol, please refer to Zhang et al. (2020).
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