We utilized WT C57BL/6 mice purchased from Envigo (Indianapolis, IN, USA). Total sixteen adult male mice (8–9 weeks, 22 ± 2 g) were housed in four ventilated Plexiglas cages (4 mice per cage). Four mice were tested in each group. Mice are maintained at temperature (21 °C) in an animal housing facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care with a reversed 12-hour dark-light cycle (lights off at 1000, lights on at 2200). This study was carried out in accordance with the recommendations of the National Institutes of Health Guide for the Care and Use of Laboratory Animals. The protocol (#1201000592 by Y.F.L. for Zebrafish, and #1605001408 by R.M.vR for mice) was approved by the Purdue University Institutional Animal Care and Use Committee.
Wt c57bl 6 mice
Manufactured by Inotiv
Sourced in United Kingdom, United States
WT C57BL/6 mice are a commonly used wild-type mouse strain. They are a standard laboratory mouse model used for a variety of research applications.
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12 protocols using wt c57bl 6 mice
1
Murine Behavior Experiments in Mice
2
Breeding C57BL/6 Mice Under SPF Conditions
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WT C57BL/6 mice were purchased from Envigo (Cambridgeshire, United Kingdom) and bred under specified pathogen-free conditions at the Epalinges Center. Five-to 10-wk-old females (to prevent potential variations as a result of sex hormones) were used for the experiments.
3
Generation of Ythdf Knockout Mouse Models
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Complete knockout models of Ythdf1 and Ythdf3 were provided by Y. Hanna (Lasman et al., 2020 (link)). GFP- and HA-tagged Ythdf2HA−GFP-Fl (DF2fl/fl) mice were provided by D. O’Carroll (Ivanova et al., 2017 (link)) and crossed to a B cell-specific Cd23-cre strain (CD23-DF2fl/fl). The Cd23-cre mice were provided by M. Busslinger (IMP, Vienna). Ythdf1−/−, Ythdf3−/− and CD23-DF2fl/fl mice were bred to mice expressing a modified NP-specific B1-8hi B cell receptor as well as a fluorescent marker (GFP or TdTomato). GFP and TdTomato-expressig mice were purchased from Jackson Laboratories. B1-8hi and B1-8lo transgenic mice were a gift from M. Nussenzweig (Rockefeller University, NY). Transgenic mice were maintained in SPF housing conditions and used at 6–18 weeks of age. WT (C57BL/6) mice were purchased from Envigo and used at 6–15 weeks of age. Both male and female mice were used throughout the study and no gender differences were observed. All experiments on mice were approved by the Weizmann Institute Animal Care and Use Committee.
4
Isolation and Culture of Murine Immune Cells
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Mice used for this study were 8–12 weeks old from a C57BL/6 background. WT C57BL/6 mice for isolation of bone marrow and generation of BMDMs and BMDCs were purchased from Envigo (Oxfordshire, United Kingdom). OT-II mice were acquired from Jackson Laboratories (Bar Harbor, United States) and breeding was maintained in house. All mice were housed in UCC Biological Services Unit under specific pathogen free (SPF) conditions. Standard housing conditions were maintained with 12 h darkness and 12 h light, temperature controlled at 21°C and 50% humidity. Animals were fed standard chow pellets and water was given ad libitum. The animal work was performed in accordance with EU legislation, in accordance with EU Directive 2010/63/EU, for the protection of animals used for scientific purposes and approved by the Animal Experimentation Ethics Committee of University College Cork, Euthanasia Only Application ID 2018/007.
5
Chronic Alcohol-Induced Liver Injury
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WT C57BL/6 mice (10–12 weeks) were purchased from Envigo Laboratories. Mice were subjected to chronic alcohol feeding to induce alcohol-induced liver injury with modification to the NIAAA model47 (link). Briefly, mice were acclimated to a liquid diet for 5 days and were fed either a 5% alcohol-containing liquid diet or an isocaloric pair-fed control diet. On day 11, IP injection of LPS (Invivogen tlrl-eblps) at 4 mg/kg was administered in alcohol-fed mice whereas PBS of the same volume was given to pair-fed mice. In LPS injection only mice, animals were fed a control diet and given LPS injection on day 11. All mice were sacrificed 8 h after LPS injection. In a subset of mice, concurrent with alcohol or pair-feeding, they were given an injection of iBET151 daily. Mice were injected intraperitoneally with iBET151 (6 mg/kg) in 10% Kleptose 2% DMSO solution or an equal volume of carrier solution. On day 11, the drug or vehicle was administered 1 h before LPS injection.
6
Alcohol-Induced Liver Injury in Mice
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WT C57BL/6 mice (10–12 weeks) were purchased from Envigo Laboratories. Mice were subjected to chronic-binge alcohol feeding to induce alcohol-induced liver injury in accordance with the NIAAA model45 (link). Briefly, mice were acclimated to a liquid diet for 5 days and were fed either a 5% alcohol-containing diet (Bio-Serv F1258SP) or an isocaloric pair-fed control diet (Bio-Serv F1259SP). All mice were gavaged with ethanol 6 g/kg (alcohol-fed mice) or an equal volume of maltose dextran (pair-fed mice) on day 11, and all mice were sacrificed 9 h later.
7
Nox1 Knockout Mouse Model
8
Murine Models of Pancreatic Cancer
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C57BL/6 mice (WT) were obtained from Envigo (Indianapolis/Indiana/USA). KC mice, KIC mice and Rag2-KO mice were bred in house (males and females). All mice were kept under specific pathogen-free conditions at the Animale en Cancérologie (AniCan) platform at the Cancer Research Center of Lyon (CRCL). All animal procedures and experiments were performed in compliance with the ethical guidelines of the CRCL Animal Care and Use Committee, with approval of the Experimental Animal Ethics Committee of the Rhône-Alpes region (CECCAPP) (CECCAPP_CLB_2019_002).
9
Transgenic Mouse Model for Immunity
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Experiments were performed according to UK Home Office regulations (PPL 30/3293, 30/2744, 30/2414) after approval by the University of Oxford ethical review board. Il21−/− transgenic knock-in mice (Jean Langhorne, National Institute for Medical Research) and C57BL/6 WT mice (Envigo, UK) were kept under conventional conditions in individually ventilated cages and fed normal chow diet.
10
Impact of Proleukin and S2A on Murine PBMCs
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C57bl/6 WT mice, 8 weeks old were obtained from Envigo and injected with 30,000 IU of Proleukin (n = 8), S2A (n = 8) and the same volume of PBS (n = 8) at day 0. At day 3 blood was drawn and PBMCs isolated for flow-cytometry (as described in the “PBMCs isolation and FACS analysis” subsection). A day later (day 4) mice were injected again in the same manner. At day 7, mice were sacrificed and blood was drawn and PBMCs isolated for flow-cytometry.
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