Male c57bl 6 mice

Male C57BL/6 mice and female BALB/c mice were purchased from Shanghai Model Organisms Center Inc. Lipopolysaccharide and TUNEL kits were purchased from Sigma, USA. In total, 10% chloral hydrate was purchased from Zhujiang Hospital of Southern Medical University. Polyvinylidene fluoride (PVDF) membranes were purchased from Millipore, USA. Protein primary antibodies were purchased from cell signaling technology, USA. Protein secondary antibody and bicinchoninic acid (BCA) kits were purchased from Shanghai Beyotime Biotechnology Co., Ltd. Enhanced chemiluminescence (ECL) kit was purchased from Bio-Rad, USA. EPO (product batch number: 20,100,901, product specification 5000 IU/branch) was purchased from China resources Angde Biotech Pharma Co., Ltd. 4% paraformaldehyde was purchased from Guangzhou Chemical Reagent Factory (GCRF). Paraffin, xylene, and neutral gum were purchased in Beijing Solarbio Science & Technology Co., Ltd. SDS-PAGE gel preparation kit and RIPA lysate were purchased from Wuhan Servicebio Co., Ltd. Malondialdehyde (MDA) and superoxide dismutase (SOD) detection kits were purchased from Nanjing Jiancheng Bioengineering Institute.

Male C57BL/6 mice (n = 190; aged 9 weeks; weighed 16‐24 g) and FOXO4 knockdown mice (as FOXO4‐KO mice; n = 20) (Shanghai Model Organisms Center Inc) were housed under a specific controlled pathogen‐free environment at the First Affiliated Hospital of Jinan University. The mice were subjected to a 12‐h light/dark cycle with controlled temperature (21°C) and humidity (60 ± 5%) and granted free access to food and water. Prior to modelling, the mice were deprived of food but not water.

All animal experiments were performed in accordance with the NIH guidelines for the Care and Use of Laboratory Animals. The study protocol was approved by the Committee on the Ethics of Animal Experiments of the Shanghai Jiaotong University School of Medicine. C57BL/6 male mice (8-12 weeks) were purchased from Shanghai Model Organisms Center, Inc. (Shanghai, China). C57BL/6 male mice were randomly assigned into two groups (vehicle group with saline and 2.5 mg/kg/d Ang II group) or four groups (vehicle group, 1.44 mg/kg/d Ang II group, 2.5 mg/kg/d Ang II group, and 2.5 mg/kg/d Ang II with valsartan group). The mice were infused with 1.44 mg/kg/d or 2.5 mg/kg/d Ang II (Sigma-Aldrich, Cat# A9525) for two weeks through subcutaneous osmotic minipumps (Model 2002, ALZA Scientific Products, Mountain View, CA, USA). These two doses of Ang II were chosen based on their well-known effects to induce vascular remodeling or abdominal aortic aneurysm and cardiac hypertrophy [17 (link)–19 (link)]. Saline-infused mice served as the controls. Valsartan (40 mg/kg/d) was administered daily via oral gavage for two weeks. The body weight and blood pressure of each mouse were measured weekly.

The care and use of animals were conducted in strict accordance with institutional guidelines and governmental regulations. All mice were maintained under a reversed 12-h /12-h day/night cycle at 22–25°C with ad libitum access to rodent food and water in environmentally controlled conditions. The mice used in the experiments were adult (8–15 weeks) C57BL/6 male mice (Shanghai Model Organisms), Vgat-ires-cre knock-in mice (Stock No. 028862) and DAT-ires-cre knock-in mice (Stock No. 006660; Jackson Laboratory, Bar Harbor, ME). The engineered mice were both maintained on a C57BL/6J genetic background.
All experiments involving mice were carried out in accordance with the US National Institutes of Health Guide for the Care and Use of Animals under an Institutional Animal Care and Use Committee approved protocol and Association for Assessment and Accreditation of Laboratory Animal Care approved Facility at the ShanghaiTech University.

Eighteen C57BL/6 male mice weighing 20–25 g were purchased from Shanghai Model Organisms Center (Shanghai, China). Hyperlipidemia was induced by the intraperitoneal administration of P-407 (P2443, Sigma-Aldrich) in PBS at a dose of 500 mg/kg three times a week. On day 30, the hyperlipidemic mice received an intraperitoneal injection of cerulein (AnaSpec, Los Angeles, CA, USA) in PBS at a dose of 50 μg/kg every hour for a total of 10 injections. Then blood samples were collected from the tail vein and centrifuged for 15 min at 3,500 g, serum was obtained for further analysis.
To evaluate the effect of hnRNPA2B1 de-neddylation in HTGP mice, MLN4924 (30 mg/kg body weight) was administered subcutaneously, twice a week for two weeks. Animals were subsequently euthanized using intraperitoneal pentobarbital (100 mg/kg; P-010, Sigma-Aldrich).

Wild-type (WT) C57BL/6 male mice aged 6–8 weeks were purchased from the Laboratory Animal Center of Chongqing Medical University, and IL-33 knockout (IL-33^−/−) mice (NM-KO-190436) on a C57BL/6 background were purchased from Shanghai Model Organisms Center (Shanghai, China). All mice were housed under specific pathogen-free conditions with free access to water and chow. After anaesthetization with an intraperitoneal injection of sodium pentobarbital (50 mg/kg), the mice were subjected to intratracheal (i.t.) instillation of 3 mg/kg lipopolysaccharide (LPS, Escherichia coli serotype O111: B4; Sigma-Aldrich, St. Louis, MO, USA) dissolved in sterile phosphate-buffered saline (PBS). A sham operation was performed in a similar manner with the same volume of PBS instead of LPS. For IL-33 injection experiments, 1000 ng of recombinant mouse IL-33 (rmIL-33) per mouse was dissolved in sterile PBS (rmIL-33, 3236-ML-010; R&D Systems) and injected intraperitoneally following intratracheal instillation of PBS [16 (link)]. Control mice received the same volume of PBS at the same time point. After 2 days of LPS insult, the mice were sacrificed for further analysis according to the Interdisciplinary Principles and Institutional Animal Care and Use Committee guidelines.