Animal experiments were approved by the UCSF Institutional Animal Care and Use Committee and were conducted in accordance with the NIH Guide for the Care and Use of Laboratory animals (protocol no. AN181214). Adult (8 to 10 weeks old) male C56BL/6 mice (strain no. 664) were purchased from the Jackson Laboratory. Mice were housed in cages on a standard 12:12 hour light-dark cycle with food and water ad libitum. The α2AAR D79N mutant mice were purchased from Jackson (stock no. 2777), and 7- to 8-week-old females were used. Sample sizes were modeled on our previous studies and on studies using a similar approach, which were able to detect significant changes (96 (link), 97 (link)). The animals were randomly assigned to treatment and control groups. Animals were initially placed into one cage and allowed to freely run for a few minutes. Then each animal was randomly picked up, injected with compound treatment or vehicle, and placed into a separate cylinder before the behavioral test.
C56bl 6 mice
Manufactured by Jackson ImmunoResearch
Sourced in United States
C56BL/6 mice are a commonly used inbred strain of laboratory mice. They are a widely utilized model organism for a variety of biomedical research applications.
Automatically generated - may contain errors
Lab products found in correlation
Sylgard 184 silicone elastomer kit, by Thermo Fisher Scientific (1 mentions)
Red cmtpx, by Thermo Fisher Scientific (1 mentions)
Green cmfda, by Thermo Fisher Scientific (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Antibodies against ampk, by Cell Signaling Technology (1 mentions)
P ampk, by Cell Signaling Technology (1 mentions)
Cisplatin, by Merck Group (1 mentions)
15 protocols using c56bl 6 mice
1
Behavioral Testing of α2A-Adrenergic Receptor Mutant Mice
2
Murine Peritoneal Adhesion Assay
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Cell-to-peritoneum adhesion was assessed ex vivo as published previously [40] (link), [41] (link). Briefly, C56BL6 mice (Jackson Laboratories) were euthanized by CO2 inhalation and subsequent cervical dislocation, then dissected using a ventral midline incision; four peritoneal tissue pieces were removed and pinned to the bottom of 24-well dishes precoated with optically transparent silicone using Sylgard 184 Silicone Elastomer Kit (Fisher). OvCa433Ncad+ and SKOV3Ecad+ cells were transiently stained with red CMTPX or green CMFDA (Invitrogen), respectively. Parental OvCa433 and SKOV3 cells were RFP- or GFP-labeled, respectively. Cells were applied to murine peritoneal explants, incubated for 1 to 2 hours, and washed with 3 × 3-minute ice-cold PBS washes, and cells/peritoneum were imaged with AMG EVOS fluorescence microscope. Image analysis was performed with ImageJ. The assay was repeated in triplicate, and statistical analysis was performed using a Student's t test. The fourth replicate tissue explants were subjected to SEM processing and imaging as described above.
3
Dosage and Toxicity Study in Mice
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Animal studies were performed according to an active Institutional Animal Care and Use Committee protocol (A377) approved by George Washington University.
Male and female C56BL/6 mice (6 to 9 weeks old) were obtained from The Jackson Laboratory. Compounds were first resuspended in DMSO at 1 M and then diluted to the appropriate concentration in Dulbecco PBS. The total DMSO concentration injected into each mouse was 4%. Five mice per group were injected with the compounds at 0, 1, 10, 50, and 100 mg/kg via the s.c. route or at 0, 1, 10, and 50 mg/kg by the i.p. route. Mice were monitored for 8 h after administration for signs of toxicity and weighed daily for 14 days. After 14 days, the mice were euthanized for necropsy.
Male and female C56BL/6 mice (6 to 9 weeks old) were obtained from The Jackson Laboratory. Compounds were first resuspended in DMSO at 1 M and then diluted to the appropriate concentration in Dulbecco PBS. The total DMSO concentration injected into each mouse was 4%. Five mice per group were injected with the compounds at 0, 1, 10, 50, and 100 mg/kg via the s.c. route or at 0, 1, 10, and 50 mg/kg by the i.p. route. Mice were monitored for 8 h after administration for signs of toxicity and weighed daily for 14 days. After 14 days, the mice were euthanized for necropsy.
4
Aging Study on C56BL/6 Mice
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Six-to-eight week old female C56BL/6 mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). Mice were maintained at the small animal facility of the National Research Council Canada (NRC) in accordance with the guidelines of the Canadian Council on Animal Care. All animals use protocols were approved by the NRC Animal Care Committee (Protocol 2011.24).
5
C56BL/6 Mouse Housing and Care
6
Mapping Neural Connections in Mice
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Adult male wild-type C56BL/6 mice (~10 wks; Jackson Laboratory, Bar Harbor, ME) were prepared similarly to methods described above. Briefly, 350 nl of Cholera Toxin subunit B (CTB) conjugated to Alexa Fluor-488, 555, or 647 (Molecular Probes, Eugene, OR, USA) was injected into the dorsal periaqueductal gray (dPAG; AP: −4.2, ML: −0.5; DV: −2.4 mm) and NAc shell (AP: +1.0, ML: +0.75, DV: −4.5 mm) (color counterbalanced between animals). Histological, imaging, and data analyses are the same as previously described.
7
C56BL/6 Mouse Husbandry Protocol
8
Murine Macrophage Response to Legionella Infection
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A/J and C56BL/6 mice were purchased from the Jackson laboratory (Bar Harbor, Maine). Caspase 1/11-/- [64 (link)] and the corresponding C56BL/6 control mice were similarly maintained. Mice lacking caspase 1, 7 and 11 were generated by intercrossing Casp1/11-/- mice [65 (link)] and Casp7-/-mice [66 (link)] (purchased from Jackson Labs, Bar Harbor, ME). The cathepsin B [52 (link)] and cathepsin H [67 (link)] knock mice (cstB-/-, cstH-/-) and the control mice were maintained in the animal facility of New York University School of Medicine. Bone marrow-derived macrophages were prepared from 6–10-week old female mice with L-supernatant as described previously [68 (link)]. Macrophages were seeded in 24-well plate the day before infection. Cell density of 4x105/well was used for intracellular bacterial growth and LDH release assay, whereas 2x105/well was used for immunofluorescence staining and other single cell-based assays. L. pneumophila used for infection was grown in AYE broth to post-exponential phase based on both optical value (OD600 = 3.4–4.0) and bacterial motility.
9
Randomized Behavioral Testing in Mice
10
Visceral Sensitivity in Aged C57BL/6 Mice
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C56BL/6 mice were obtained from Jackson lab and bred at the University of South Carolina. Male and female C57BL/6 mice were used, due to their visceral sensitivity response to colorectal distension [52 (link)]. A conditioned place aversion test was performed at 24–25 months of age. Body weight was measured, and blood collected for complete blood count.
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