Evaluation of mTORC1 signaling in mouse atherosclerotic macrophages by flow cytometry and immunofluorescence

Xiangyu Zhang; Jeremiah Stitham; Astrid Rodriguez-Velez; Se Jin Jeong; Arick Park; Yu Sheng Yeh; Divya Kapoor; Bettina Mittendorfer; Babak Razani

doi:10.1016/j.xpro.2022.101665

Evaluation of mTORC1 signaling in mouse atherosclerotic macrophages by flow cytometry and immunofluorescence

Xiangyu Zhang, Jeremiah Stitham, Astrid Rodriguez-Velez, Se Jin Jeong, Arick Park, Yu Sheng Yeh, Divya Kapoor, Bettina Mittendorfer, Babak Razani

Research output: Contribution to journal › Article › peer-review

Abstract

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 atherosclerosis-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).

Original language	English
Article number	101665
Journal	STAR Protocols
Volume	3
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2022.101665
State	Published - Dec 16 2022

Keywords

Cell isolation
Flow cytometry/Mass cytometry
Immunology

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10.1016/j.xpro.2022.101665

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title = "Evaluation of mTORC1 signaling in mouse atherosclerotic macrophages by flow cytometry and immunofluorescence",

abstract = "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 atherosclerosis-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).",

keywords = "Cell isolation, Flow cytometry/Mass cytometry, Immunology",

author = "Xiangyu Zhang and Jeremiah Stitham and Astrid Rodriguez-Velez and Jeong, {Se Jin} and Arick Park and Yeh, {Yu Sheng} and Divya Kapoor and Bettina Mittendorfer and Babak Razani",

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T1 - Evaluation of mTORC1 signaling in mouse atherosclerotic macrophages by flow cytometry and immunofluorescence

AU - Zhang, Xiangyu

AU - Stitham, Jeremiah

AU - Rodriguez-Velez, Astrid

AU - Jeong, Se Jin

AU - Park, Arick

AU - Yeh, Yu Sheng

AU - Kapoor, Divya

AU - Mittendorfer, Bettina

AU - Razani, Babak

PY - 2022/12/16

Y1 - 2022/12/16

N2 - 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 atherosclerosis-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).

AB - 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 atherosclerosis-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).

KW - Cell isolation

KW - Flow cytometry/Mass cytometry

KW - Immunology

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DO - 10.1016/j.xpro.2022.101665

M3 - Article

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AN - SCOPUS:85137643122

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VL - 3

JO - STAR Protocols

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Evaluation of mTORC1 signaling in mouse atherosclerotic macrophages by flow cytometry and immunofluorescence

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Keywords

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