B6.129 cg ccr2tm2 1ifc j

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B6.129(Cg)-Ccr2tm2.1Ifc/J is a transgenic mouse strain carrying a targeted mutation in the Ccr2 gene. The Ccr2 gene encodes the C-C chemokine receptor type 2, which is involved in the migration and function of monocytes and macrophages. This mouse strain can be used in research applications that require the study of the Ccr2 signaling pathway and its role in various biological processes.

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12 protocols using b6.129 cg ccr2tm2 1ifc j

Generation and Characterization of Genetically Modified Mice

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Wild-type C57BL/6, Ccr2^RFP reporter (B6.129(Cg)-Ccr2^tm2.1Ifc/J), Cx3cr1^GFP reporter (B6.129P-Cx3cr1^tm1Litt/J), CD45.1 (B6.SJL-Ptprc^a Pepc^b/BoyJ), Raptor ^fl/fl (B6.Cg-Rptor^tm1.1Dmsa/J), ubiquitin-ERT2-cre (B6.Cg-Tg(UBC-cre/ERT2)1Ejb/1J), Mx1-Cre (B6.Cg-Tg(Mx1-cre)1Cgn/J), and LysM-Cre (B6.129P2-Lyz2^tm1(cre)Ifo/J) mice were obtained from Jackson Laboratories. Femurs from Myc^GFP mice (B6;129-Myc^tm1Slek/J) were also obtained from Jackson Laboratories. Ccr2^RFP/+ Cx3cr1^GFP/+ (dual reporter) mice, Cre/ERT2 Raptor ^fl/fl mice (Raptor iKO), Mx1-Cre Raptor ^fl/fl mice and LysMCre Raptor ^fl/fl mice were generated as described (19 (link), 26 (link), 56 (link)). Tsc2 ^fl/fl (Tsc2^tm1.1Mjg/J) mice were kindly provided by Dr. Michael Gambello (57 (link)) and bred with Mx1-Cre mice to generate Tsc2 KO mice. Male and female mice age between 6 to 8 weeks were used with matched littermate controls.

Lee P.Y., Sykes D.B., Ameri S., Kalaitzidis D., Charles J.F., Nelson-Maney N., Wei K., Cunin P., Morris A., Cardona A.E., Root D.E., Scadden D.T, & Nigrovic P.A. (2017). The metabolic regulator mTORC1 controls terminal myeloid differentiation. Science immunology, 2(11), eaam6641.

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Multimodal Immune Cell Profiling

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C57BL/6J, B6.129(Cg)-Ccr2^tm2.1Ifc/J (CCR2-RFP), B6.Cg-Tg(Itgax-Venus)1Mnz/J (CD11c-YFP), B6(Cg)-Tyr^c−2J/J (B6 albino), B6.Cg-Zbtb46^{tm4.1(HBEGF)MnzTyrc−2J}/J (Zbtb46-DTR), B6.129P2(C)-Ccr7^tm1Rfor/J (CCR7.KO), B6.129S1-Il12^btm1Jm/J (IL-12p40.KO), and B6.129S4-Ccr2^tm1Ifc/J (CCR2.KO) mouse strains were obtained from The Jackson Laboratory. CD45.1⁺ B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II), CD45.1⁺ C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I), and B6.SJL-Ptprc^aPepc^b/BoyCrl (CD45.1⁺) were obtained either from donating investigators (Dr. Pamela J. Fink, University of Washington) or Charles River. CD11c-YFP animals were crossed with B6 albino mice to homozygosity, and next crossed to CCR2-RFP mice to generate a CD11c-YFP x CCR2-RFP^Hetrozygous dual reporter mice. CCR2-DTR mice were obtained from donating investigators (Dr. Steven F. Ziegler, Benaroya Research Institute) and with approval from the originating investigators (Drs. Tobias M. Hohl and Eric G. Pamer, Memorial Sloan-Kettering Cancer Center) (95 (link)). 6–10 week-old male and female mice were kept in specific pathogen–free conditions at an Association for Assessment and Accreditation of Laboratory Animal Care–accredited animal facility at the University of Washington, South Lake Union campus. All procedures were approved by the University of Washington Institutional Animal Care and Use Committee.

Leal J.M., Huang J.Y., Kohli K., Stoltzfus C., Lyons-Cohen M.R., Olin B.E., Gale M J.r, & Gerner M.Y. (2021). Innate cell microenvironments in lymph nodes shape the generation of T cell responses during Type-I inflammation. Science immunology, 6(56), eabb9435.

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Obesity-Induced Wound Healing in CCR2-RFP Mice

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Homozygous B6.129(Cg)-Ccr2^tm2.1Ifc/J mice and C57Bl/6 were purchased from the Jackson Laboratory (Bar Harbor, ME, USA) and crossbred to generate heterozygous B6.129(Cg)-Ccr2^tm2.1Ifc/J (CCR2^RFP/+) mice in which CCR2+ cells express red fluorescent protein (RFP) to allow tracking of these cells. For the induction of diet-induced obesity, male and female CCR2^RFP/+ mice (n = 15–17/group/sex) were fed a high-fat diet (HFD, 60 Kcal% fat, 7% Kcal/fructose, Research Diets) ad libitum for 12 weeks, beginning at 8 weeks of age; mice were thus ~20 weeks old at the time of wounding. Age-matched lean CCR2^RFP/+ mice received a standard chow diet (ND; n = 15–17/group/sex). All mice were housed in environmentally controlled conditions with a 12 h light/dark cycle. Water and food were available ad libitum. All animal studies were approved by the Animal Care and Use Committee of the University of Illinois at Chicago.

Pang J., Urao N, & Koh T.J. (2024). Diet-Induced Obesity Increases Monocyte/Macrophage Proliferation during Skin Wound Healing in Mice. Cells, 13(5), 401.

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Generating Transgenic GFP/RFP Mice for Microglial and Monocyte Tracking

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C57BL/10SnJ mice were obtained from an in-house breeding colony, these mice are noted as C57BL/10 throughout the paper. TgGFP/RFP mice were generated by crossbreeding Cx3cr1 knockout mice homozygous for the Cx3cr1-GFP targeted mutation (B6.129P-Cx3cr1^tm1Litt/J,The Jackson Laboratory, Stock No: 005582) with Ccr2 knockout mice homozygous for the Ccr2-RFP targeted mutation (B6.129(Cg)-Ccr2^tm2.1Ifc/J,The Jackson Laboratory, Stock No:017586) [21 (link), 34 (link)]. Resultant offspring had heterozygous expression of both Cx3cr1 and Ccr2. In addition, green fluorescent protein (GFP) was expressed under promoter for Cx3cr1, prominently expressed by microglia in the CNS and red fluorescent protein (RFP) was expressed under the promoter for Ccr2, prominently expressed by monocytes [33 (link)]. All mice were group housed in transparent cages in a 12 h light (250-300 lx) /12 h dark cycle and food and water were available ad libitium.

Striebel J.F., Race B., Williams K., Carroll J.A., Klingeborn M, & Chesebro B. (2019). Microglia are not required for prion-induced retinal photoreceptor degeneration. Acta Neuropathologica Communications, 7, 48.

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Breeding and Maintenance of CCR2 Mice

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Female wild-type C57BL/6JRj mice were purchased from Janvier Labs. CCR2-deficient (B6.129S4-Ccr2^tm1Ifc/J), CCR2^RFP [B6.129(Cg)-Ccr2^tm2.1Ifc/J], and C57BL/6NCrl mice were bred and maintained at the Lund University Clinical Research Center (CRC), Malmö. B6.129S4-Ccr2^tm1Ifc/J and B6.129(Cg)-Ccr2^tm2.1Ifc/J were originally purchased from The Jackson Laboratory. B6.129S4-Ccr2^tm1Ifc/J were crossed with C57BL/6NCrl mice, originally obtained from Charles River lab, to generate CCR2^+/− mice for CCR2^+/− × CCR2^−/− breeding. Littermate controls were used for experiments involving CCR2^+/− and CCR2^−/− mice.

Lienard J., Munke K., Wulff L., Da Silva C., Vandamme J., Laschanzky K., Joeris T., Agace W, & Carlsson F. (2023). Intragranuloma Accumulation and Inflammatory Differentiation of Neutrophils Underlie Mycobacterial ESX-1-Dependent Immunopathology. mBio, 14(2), e02764-22.

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Generating CCR2-deficient Cilia Mutant Mice

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Male and female, 8-week-old, CAGG-Cre/Esr1/5Amc/J Ift88^f/f (referred to as conditional Ift88 mice) C57BL/6J mice were bred in house. CCR2^rfp/rfp mice (B6.129[Cg]-Ccr2tm2.1Ifc/J; stock number 017586) were purchased from The Jackson Laboratories and crossed to the conditional Ift88 mice to generate the CCR2-deficient, cilia mutant mice. Animals were maintained in facilities accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International, in accordance with the Institutional Animal Care and Use Committee regulations at the University of Alabama at Birmingham (UAB; approval numbers, 10130 and 21072).

Aloria E.J., Song C.J., Li Z., Croyle M.J., Mrug M., Zimmerman K.A, & Yoder B.K. (2021). Ly6chi Infiltrating Macrophages Promote Cyst Progression in Injured Conditional Ift88 Mice. Kidney360, 2(6), 989-995.

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Investigating Immune Cell Dynamics

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Male and female C57BL/6 (#000664 Jackson Laboratories), Ccr2 reporter knock in (B6.129(Cg)-Ccr2tm2.1Ifc/J, #017586 Jackson Laboratories) Tcrb^−/− (B6.129P2-Tcrbtm1Mom/J, #002118 Jackson Laboratories), and Cd4^−/− mice (B6.129S2-Cd4tm1Mak/J, #002663Jackson laboratories) were used for these studies and maintained on a congenic background. All researches conducted on animals were approved by the Institutional Animal Care and Use Committee at the University of Alabama at Birmingham.

Williams G.P., Marmion D.J., Schonhoff A.M., Jurkuvenaite A., Won W.J., Standaert D.G., Kordower J.H, & Harms A.S. (2020). T cell infiltration in both human multiple system atrophy and a novel mouse model of the disease. Acta Neuropathologica, 139(5), 855-874.

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Genetically Modified Mouse Strains

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C57BL/6, B6.Cg‐Kit^W‐sh/HNihrJaeBsmJ (Kit^{W‐sh/W‐sh}), B6.129‐Tlr2^tm1Kir/J (Tlr2^–/–), B6.129S1‐Nod2^tm1Flv/J (Nod2^–/–), B6.129(Cg)‐Ccr2^tm2.1Ifc/J (Ccr2^–/–) and B6.129S1‐Ccr5^tm1Kuz/J (Ccr5^–/–) were purchased from The Jackson Laboratory (Bar Harbor, ME) and bred in‐house. CXCR3‐deficient animals (Cxcr3^–/–) were generously provided by Dr Brent Johnston (Dalhousie University, Halifax, Canada) and bred in‐house. CCR4‐deficient animals (Ccr4^–/–) were generously provided by Dr Steven Kunkel (University of Michigan, Ann Arbor, Michigan) and bred in‐house. Myd88 knockout animals were generously provided by Dr S. Akira (Osaka University, Osaka, Japan) and bred in‐house. Nod2 and Tlr2 knockouts were bred to produce double knockouts for both Nod2 and Tlr2. Female mice aged 7–14 weeks were used in all experiments. Mice were housed under specific pathogen‐free conditions with food and water provided ad libitum. All experiments followed the guidelines provided by the Canadian Council on Animal Care and were performed according to protocols approved by the Animal Research Ethics Board of Dalhousie University (Halifax, Canada).

Haidl I.D., Meghnem D., Issekutz T.B, & Marshall J.S. (2020). Toll‐like receptor 2 activation induces C–C chemokine receptor 2‐dependent natural killer cell recruitment to the peritoneum. Immunology and Cell Biology, 98(10), 854-867.

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Breeding and Characterization of Immune Knockout Mice

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All mice were bred and housed under SPF conditions in the Carlson Barrier Facility at the University of Chicago. Wild-type C57BL/6J mice (Stock 000664), Ccl2^−/− mice (B6.129S4-Ccl2tm1Rol/J, Stock 004434), Ccr2^RFP/RFP mice (B6.129(Cg)-Ccr2tm2.1Ifc/J, Stock 017586), and CD45.1⁺ mice (B6.SJL-Ptprca Pepcb/BoyJ, Stock 002014), were purchased from The Jackson Laboratory. Ccr4^−/− mice were kindly provided by Dr. John Belperio (University of California Los Angeles).

Chang A.L., Miska J., Wainwright D.A., Dey M., Rivetta C.V., Yu D., Kanojia D., Pituch K.C., Qiao J., Pytel P., Han Y., Wu M., Zhang L., Horbinski C.M., Ahmed A.U, & Lesniak M.S. (2016). CCL2 produced by the glioma microenvironment is essential for the recruitment of regulatory T cells and myeloid-derived suppressor cells. Cancer research, 76(19), 5671-5682.

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CCR2+ Monocyte Tracking in Inflammation

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CCR2-RFP reporter mice were purchased from The Jackson Laboratory (B6.129(Cg)-Ccr2^tm2.1Ifc/J). These mice have a monomeric RFP sequence replacing the coding sequence of the chemokine (C-C motif) receptor 2 (Ccr2) gene and is useful to track CCR2⁺ monocyte recruitment to sites of inflammation.

Arnold T.D., Lizama C.O., Cautivo K.M., Santander N., Lin L., Qiu H., Huang E.J., Liu C., Mukouyama Y.S., Reichardt L.F., Zovein A.C, & Sheppard D. (2019). Impaired αVβ8 and TGFβ signaling lead to microglial dysmaturation and neuromotor dysfunction. The Journal of Experimental Medicine, 216(4), 900-915.

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