8–10 week old female wild-type (WT) and Gadd45b knockout (KO) mice littermates with C57BL/6 background were used (Charles River). Sample sizes and grouping were based on previous study showing significant differences following ECS [13 (link)]. Animals received a single ECS via ear-clip electrodes using the Ugo Basile ECS unit (Model 7801) as previously described [13 (link),14 (link)]. Under the specific parameters (1.0 s, 100 Hz, 18 mA stimulus of 0.3 ms square wave pulses), animals initially exhibited full hind limb extension for 1–2s with an overall seizure duration of > 20s and full recovery in < 5 min [13 (link),14 (link)]. Both Gadd45b WT and KO mice showed similar convulsions. Sham animals were similarly handled in parallel without ECS. Three days later, mice were sacrificed, fixed, and brains subjected to analysis. Previous studies show a single ECS treatment induces significant proliferative effects visible at this time point [5 (link),13 (link)]. All experimental procedures were performed in accordance with the animal protocol approved by the Institutional Animal Care and Use Committee.
C57bl 6 background
Manufactured by Charles River Laboratories
Sourced in China
The C57BL/6 background is a mouse strain commonly used in biomedical research. It is a widely utilized genetic background for the creation of genetically modified mouse models. The C57BL/6 background provides a standardized foundation for research studies.
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Lab products found in correlation
B6.129 gt rosa 26sortm1 cre ert2 tyj j, by Jackson ImmunoResearch (1 mentions)
Cd45.1 mice, by Charles River Laboratories (1 mentions)
C57bl 6 background, by Jackson ImmunoResearch (1 mentions)
Cd11c promoter, by Jackson ImmunoResearch (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Hyaluronidase 4 s, by Merck Group (1 mentions)
Tyrode s, by Merck Group (1 mentions)
Bgc823, by National Collection of Authenticated Cell Cultures (1 mentions)
Mgc 803, by National Collection of Authenticated Cell Cultures (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin plus streptomycin, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
12 protocols using c57bl 6 background
1
Electroconvulsive Seizure in Mice
2
SFG Intravenous Injection in Mice
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Eight-week-old male mice with C57BL/6 background were obtained from Charles River Laboratories, and all animals were hosted in single ventilated cages in a 12-h day/night cycle under specific pathogen-free conditions. This animal experiment was approved by the animal ethics committee in Baden-Württemberg. SFG was diluted in sterile saline, and three mice were intravenously injected with a dosage of 625 μg in 100 μl saline. Three control animals received the same volume of saline. All mice were sacrificed after 12 h. Blood parameters and serum iron were measured by a scil Vet abc Plus device (scil animal care company GmbH).
3
Genetically Modified Mouse Protocols
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Studies were performed in accordance with protocols approved by the Animal Studies Committee of Drexel University College of Medicine in accordance with the National Institutes of Health Guidance for the Care and Use of Laboratory Animals. All experiments were performed using 8‐ to 12‐week‐old female mice that have been backcrossed extensively into the C57BL/6 background (Charles River). Generation of Rgs2+/− and Rgs2−/− mice has been described previously (Oliveira‐Dos‐Santos et al. 2000 ). Mice were provided access to food and water ad libitum in our institution's animal facility at 22°C and a 12‐h light/dark cycle.
4
Atrium-specific AMPK α1 α2 dKO Mice
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We generated mice with an atrium-specific AMPK α1 α2 dKO by initially crossing Prkaa1fl/fl and Prkaa2fl/fl mice (47 (link)) on a C57BL/6 background (Charles River). Then, homozygous Prkaa1fl/flPrkaa2fl/fl mice were crossed with C57BL/6 mice (48 (link)) expressing Cre-recombinase under the control of the Sln promoter (Sln-Cre+/–) to drive transcription in the atria. Mice expressing the Sln Cre-recombinase alone (Sln-Cre+/– Prkaa1+/+ Prkaa2+/+) were used as additional controls. Mice were backcrossed in the C57BL/6 background for at least 7 generations.
After completion of these studies, this colony was relocated, and genotyping was performed by Transnetyx using RT-qPCR with primers to WT, floxed, and excised Prkaa1 and Prkaa2 alleles (Supplemental Table 2 ).
After completion of these studies, this colony was relocated, and genotyping was performed by Transnetyx using RT-qPCR with primers to WT, floxed, and excised Prkaa1 and Prkaa2 alleles (
5
Conditional Knockdown of Decorin and Biglycan
6
Murine Model of Langerhans Cell Histiocytosis
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All animal experiments performed in this study were approved by the Institutional Animal Care and Use Committee of Mount Sinai School of Medicine. BRAFV600ECD11c mice were created by crossing BRAFV600Eca/wt or BRAFV600Eca/ca mice (provided by M.W. Bosenberg, Yale University, New Haven, CT; Dankort et al., 2007 (link)) with mice expressing cre recombinase under the control of the CD11c promoter (C57BL/6 background; The Jackson Laboratory). BRAFV600ECD11c mice with one BRAFV600ECA allele and one CD11c-Cre allele develop a lethal LCH-like syndrome, as described (Berres et al., 2014 (link)). All animals were housed under specific pathogen-free conditions and sacrificed at the indicated time points. All experiments were controlled using littermates negative for the cre recombinase transgene construct or cre-positive littermates negative for the BRAFV600Eca construct.
BRAFV600ECD11c chimeras were generated by transplantation of 1–3 × 10^6 whole BM cells flushed from the long bones of 5–8 wk old BRAFV600ECD11c mice into lethally irradiated CD45.1 mice age 8–12 wk (C57BL/6 background; Charles River Laboratory). Mice were allowed to recover for 3 wk after transplantation before initiation of drug treatments.
BRAFV600ECD11c chimeras were generated by transplantation of 1–3 × 10^6 whole BM cells flushed from the long bones of 5–8 wk old BRAFV600ECD11c mice into lethally irradiated CD45.1 mice age 8–12 wk (C57BL/6 background; Charles River Laboratory). Mice were allowed to recover for 3 wk after transplantation before initiation of drug treatments.
7
Cardiovascular Study in C57BL/6 Mice
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Male mice aged 8 to 9 weeks were used in this study (C57BL/6 background, Charles River Laboratories). Animal studies were approved by the Centro Nacional de Investigaciones Cardiovasculares (CNIC) ethics committee and conformed to directive 2010/63EU and recommendation 2007/526/EC regarding the protection of animals used for experimental and other scientific purposes, enforced in Spanish law under RD1201/2005.
8
Isolation and Preparation of Mouse Oocytes
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oocytes were obtained 6–8 weeks old wildtype female mice (C57Bl6 background, Charles River Laboratories, USA). Female mice were super-ovulated by intraperitoneal injections, first of 5 IU PMSG, followed by 5 IU hCG 48 hours apart. Cumulus-intact oocytes were collected into a Ferticult® medium drop 14 hours later by tearing the oviduct ampulla from sacrificed mice. Oocytes were separated from their cumulus by a brief incubation at 37 °C, in presence of hyaluronidase IV-S® (Sigma-Aldrich, USA) (15 mg.mL-1). Oocytes at metaphase II stage were selected on the basis of the presence of the first polar body. The Zona Pellucida (ZP) was subsequently removed by rapid treatment (<30 sec.) of the eggs with acidic Tyrode’s® (Sigma-Aldrich, USA) solution. Eggs were then incubated in Ferticult® for 2 hours at 37 °C, 5% CO2 in air to recover after the treatment. Oocytes were then incubated with Hoechst 33342 (Sigma-Aldrich, USA) at 1 μg.ml−1 for 5 minutes and washed.
9
P2Y6 Receptor Knockout Mice Protocol
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All animal work was carried out in accordance with the Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012 following ethical review by the University of Cambridge Animal Welfare and Ethical Review Body (AWERB). P2Y6R knockout mice were kindly provided by Bernard Robaye (ULB Brussels) and maintained on a C57BL/6 background (Charles River Laboratories). P2Y6R knockout mice and wild‐type littermates were used to establish homozygous P2Y6R wild‐type and knockout sub‐lines.
10
Genetic Tools for Neuroscience Research
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Rorβ-IRES2-Cre-DRorβCre/+; Jackson Laboratories (JAX), no. 023526 (20 (link)), Sst-IRES-Cre (SstCre/+; JAX, no. 028864) (58 (link)), Pv-IRES-Cre (PVCre/+; JAX, no. 017320) (59 (link)), R26:lacZbpA(flox)DTA (DTAf/+) (60 (link)), Ai14(RCL-TdT)-D (Tomatof/+; JAX, no. 007914) (61 (link)), and R26-tm1(CAG-EGFP)Fsh (GFPf/+; JAX, no. 032038) (62 (link)) were maintained in a C57BL/6 background (Charles River Laboratories). The morning of the appearance of a vaginal plug was defined as E0.5. Animals were maintained under standard, temperature-controlled conditions on a 12-hour:-12-hour light/dark cycle. Water and food were provided ad libitum. Animals were housed and maintained following the guidelines from the European Union Council Directive (86/609/ European Economic Community). All the procedures for handling and sacrificing animals followed the European Commission guidelines (2010/63/EU). All animal procedures were approved by the Consejo Superior de Investigaciones Científicas (CSIC) and the Community of Madrid Ethics Committees on Animal Experimentation in compliance with national and European legislation (PROEX 118-14; 233/16; 124-17; 234-16; 123-17; 065/19; 162.6/23; 2020/VSC/PEA/0227).
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