GFP+ donor mice (C57BL/6-Tg(CAG-EGFP) were purchased from Jackson Laboratory (Bar Harbor, ME)(Jax stock #006567). Briefly, a single cell suspension from GFP+ donor mice was prepared from the spleens of donor mice with a 40μm strainer, lymphocyte/monocyte populations were purified using separation medium and one million cells were injected per mouse via the tail vein. GFP+ cells were adoptively transferred into naïve or EAP mice 7 days after induction of prostatitis. 3 days post-adoptive transfer of GFP-positive cells behavioral testing was performed and sacral spinal cords were isolated. GFP+ cells were enumerated by flow cytometry and immunophenotyped.
C57bl 6 tg cag egfp
Manufactured by Jackson ImmunoResearch
Sourced in Montenegro
C57BL/6-Tg(CAG-EGFP) is a genetically modified mouse strain that expresses enhanced green fluorescent protein (EGFP) under the control of the CAG promoter. This transgenic mouse line is commonly used as a source of EGFP-expressing cells and tissues for research and experimental purposes.
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9 protocols using c57bl 6 tg cag egfp
1
GFP+ Cell Transfer in Prostatitis
2
STZ-Induced Diabetic Mouse Model
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All procedures involving the animal models were approved by the Institutional Animal Care and Use Committee at Michigan State University. Male C57BL/6J and C57BL/6-Tg(CAG-EGFP) mice were purchased from Jackson Laboratory and made diabetic by injections of streptozotocin (STZ) dissolved in 0.5% sodium citrate buffer, with a daily dose of 65mg/Kg for five consecutive days. The control mice received sodium citrate buffer only. Mice with blood glucose greater than 13.8 mmol/L were considered diabetic. Starting 14 days after STZ injections, insulin injections (with a dose of 0–2 units/day) were administered to prevent acute weight loss, but allowing hyperglycemia in the range of 20 mmol/L blood glucose.
3
Neutrophil Migration Assay with TMEV Stimuli
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Peripheral blood cells from naïve green fluorescent protein (GFP)-expressing C57BL/6 Tg mice [C57BL/6 Tg(CAG-EGFP), Jackson Laboratories, Maine] were used as the source of neutrophils. After removal of red blood cells using red blood cell lysing buffer (R7757, Sigma Aldrich), the cells were further purified using negative magnetic cell sorting, which utilizes the antibodies for CD2, CD5, CD45R, F4/80, and ICAM-1, as previously described (Cotter et al., 2001 (link); Yona et al., 2010 (link)). A standard micropore filter assay using Boyden chambers was performed (Gho et al., 1999 (link)). Briefly, the lower well of the chamber of a 12-well plate was filled with conditioned media from BHK monolayers mock-infected and infected with TMEV, KC-TMEV, or HEL-TMEV for 24 h at 10 MOI. The upper chambers containing transwells (8 μm pore size, Corning, #353182) were placed on the conditioned media. The isolated neutrophils were added to the transwells and incubated in 5% CO2 for 3 h at 37°C. The number of cells on the outside of the transwell membrane were enumerated using fluorescence microscopy.
4
Streptozotocin-Induced Diabetes in Mice
5
Bone Marrow Transplant Chimera Creation
6
Diabetic Retinopathy in Transgenic Mice
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All procedures involving animal models were approved by Institutional Animal Care and Use Committee at MSU. The Tie-2 promoter driven miR-15a transgenic (Tie2-miR-15a TG) mice were from Dr. Y. Eugene Chen's group (Yin et al., 2012 (link)). C57BL/6J and C57BL/6-Tg(CAG-EGFP) mice were purchased from Jackson Laboratory. Eight week old male Tie2-miR-15a TG, C57BL/6J or C57BL/6-Tg(CAG-EGFP) mice were made diabetic by injections of 55 mg STZ (Sigma-Aldrich)/kg body weight for five consecutive days. Eight week old male Long Evans rats with body weights of 240 g were purchased from the Harlan laboratories (Haslett, MI, USA) and made diabetic by injections of 65 mg STZ (Sigma-Aldrich)/kg body weight for five consecutive days.
Control animals (mice and rats) received vehicle (100 mM citric acid buffer, pH = 4.5) injections. Body weight and blood glucose were monitored biweekly for these mice and rats during the induction of diabetes. Permeability was examined 4 weeks after the induction of diabetes to mimic early stage diabetic retinopathy for the control and STZ- induced Tie2-miR-15a TG mice. Sample and data were collected 8 weeks after the induction of diabetes for real-time PCR analysis and CACs isolation for the STZ- induced Tie2-miR-15a TG mice, C57BL/6J, C57BL/6-Tg(CAG-EGFP) mice and rats.
Control animals (mice and rats) received vehicle (100 mM citric acid buffer, pH = 4.5) injections. Body weight and blood glucose were monitored biweekly for these mice and rats during the induction of diabetes. Permeability was examined 4 weeks after the induction of diabetes to mimic early stage diabetic retinopathy for the control and STZ- induced Tie2-miR-15a TG mice. Sample and data were collected 8 weeks after the induction of diabetes for real-time PCR analysis and CACs isolation for the STZ- induced Tie2-miR-15a TG mice, C57BL/6J, C57BL/6-Tg(CAG-EGFP) mice and rats.
7
Transgenic Mouse Strains for Immunology
8
Renal Ischemia-Reperfusion Injury in Mice
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C57BL/6-CCR2−/− and C57BL/6-Tg (CAG-EGFP) mice were purchased from Jackson Laboratory, USA. The control for C57BL/6-CCR2−/− mice were wild type littermates from heterozygous parents. NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju (NCG) mice, the genetic background of which are NOD/ShiLtJNju, were purchased from Nanjing Model Animal Research Center (Nanjing, China). All mice were maintained and housed in accordance with the Experimental Animal Ethics Committee of Huazhong University of Science and Technology. Mice were anaesthetized with 1% sodium pentobarbital solution (0.01 mL/g body weight, Sigma, USA) by intraperitoneal injection. The left renal pedicle was clamped with an atraumatic vascular clip for 30 min (Roboz Surgical Instrument Co, Germany) through a flank incision and the left kidney turned black subsequent to clamping. Clamps were removed after 30 min. Body temperature was controlled at 36.8–37.2 °C throughout the procedure (FHC, USA). Sham animals were subjected to a similar surgical procedure without clamping the left kidney pedicle.
9
Breeding and housing Hem1 knockout mice
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Hem1 knockout mice were bred in our facility from pairs of mice heterozygous for the Hem1 mutant allele (C57BL/6 background), as described previously (13 (link)). C57BL/6-Tg(CAG-EGFP) mice were purchased from Jackson Laboratories (Bar Harbor). All mice used in this study were housed under standard laboratory conditions with a 12 h dark, 12 h light cycle, a constant temperature of 23 °C, and humidity of 48%. A standard rodent diet (Envigo, Teklad 22/5) containing 22% protein, 1.13% calcium, and 0.94% phosphorus was provided to mice ad libitum. All mice were maintained at the University of Arkansas for Medical Sciences animal facility, which is accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care International. Mice were randomly assigned to four or five mice per cage. All animal procedures were approved by the University of Arkansas for Medical Sciences Institutional Animal Care and Use Committee.
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