Protocol for PPP1R15A-inhibited mouse model establishment with subcutaneous B16F1 tumor and single-cell analysis

Rongjing Wang; Minghui Wang; Siyu Pei; Yuchao Zhang; Shiwei Guo; Wei Guo; Zhenchuan Wu; Hailong Wang; Yizhe Li; Yufei Zhu; Ling-Hua Meng; Jingyu Lang; Gang Jin; Yichuan Xiao; Landian Hu; Xiangyin Kong

STAR Protocols (Dec 2023)

Protocol for PPP1R15A-inhibited mouse model establishment with subcutaneous B16F1 tumor and single-cell analysis

Rongjing Wang,
Minghui Wang,
Siyu Pei,
Yuchao Zhang,
Shiwei Guo,
Wei Guo,
Zhenchuan Wu,
Hailong Wang,
Yizhe Li,
Yufei Zhu,
Ling-Hua Meng,
Jingyu Lang,
Gang Jin,
Yichuan Xiao,
Landian Hu,
Xiangyin Kong

Affiliations

Rongjing Wang: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China
Minghui Wang: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; University of Chinese Academy of Sciences, Shanghai, China
Siyu Pei: Department of Thoracic Surgical Oncology, Shanghai Lung Cancer Center, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200030, China
Yuchao Zhang: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China
Shiwei Guo: Changhai Hospital, Department of Hepatobiliary Pancreatic Surgery, Shanghai, China
Wei Guo: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China
Zhenchuan Wu: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; University of Chinese Academy of Sciences, Shanghai, China
Hailong Wang: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; ShanghaiTech University, School of Life Science and Technology, Shanghai, China
Yizhe Li: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; University of Chinese Academy of Sciences, Shanghai, China
Yufei Zhu: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China
Ling-Hua Meng: Division of Anti-tumor Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China
Jingyu Lang: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China
Gang Jin: Changhai Hospital, Department of Hepatobiliary Pancreatic Surgery, Shanghai, China; Corresponding author
Yichuan Xiao: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; Corresponding author
Landian Hu: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; Anda Biology Medicine Development (Shenzhen) Co., Ltd, Shenzhen, China; Corresponding author
Xiangyin Kong: Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai, China; ShanghaiTech University, School of Life Science and Technology, Shanghai, China; Corresponding author

Journal volume & issue: Vol. 4, no. 4
p. 102616

Abstract

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Summary: Here, we present a protocol for exploring the effects of PPP1R15A inhibitor, Sephin1, on antitumor immunity of B16F1 subcutaneous tumor in mice. We describe steps for constructing single-cell transcriptome and TCR libraries, sequencing, and using sequencing data for the integration of expression and TCR data. We then detail procedures for gene differentiation, regulon and cell-cell communication analysis, and validation of single-cell analysis results.For complete details on the use and execution of this protocol, please refer to Wang et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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