Wild type C57BL/6 (Jackson # 000664), Casp1–/– (Rauch et al., 2017 (link)), Casp11–/– (Kayagaki et al., 2011 (link)), Casp1-11DKO (Kuida et al., 1995 (link)), Nlrc4-AscDKO (Mariathasan et al., 2004 (link)), Gsdmd–/– (Rauch et al., 2017 (link)), 129/SeVe (Jackson # 002448), Mrp8-Cre (Jackson # 021614), and Casp11fl/fl (Cheng et al., 2017 (link); Lee et al., 2018 (link)) mice were used in this study. All mice used in this study, except 129/SeVe, were in C57BL/6 genetic background and were bred and maintained in a specific pathogen-free facility at the University of Arkansas for Medical Sciences. For the deletion of caspase-11 in neutrophils, Casp11fl/fl mice were crossed with Mrp8-Cre. Mice which were heterozygous for the floxed gene and hemizygous for the Mrp8-Cre were backcrossed to mice homozygous for the floxed gene. Both female and male mice aged between 6 and 12 weeks were randomly subjected to the experiments and there was no difference in experimental results due to sex differences. All protocols met the guidelines of the US National Institutes of Health for the humane care of animals and were approved by the Institutional Animal Care and Use Committee at the University of Arkansas for Medical Sciences.
Mrp8 cre
Manufactured by Jackson ImmunoResearch
Sourced in Singapore
Mrp8-Cre is a Cre recombinase transgenic mouse line that expresses Cre recombinase under the control of the myeloid-related protein 8 (Mrp8) promoter. Mrp8-Cre allows for the targeted deletion of floxed genes in myeloid cells, such as macrophages and neutrophils.
Automatically generated - may contain errors
Lab products found in correlation
C57bl 6, by Jackson ImmunoResearch (3 mentions)
Lyz2 cre, by Jackson ImmunoResearch (3 mentions)
C57bl 6j, by Jackson ImmunoResearch (3 mentions)
Lysm cre, by Jackson ImmunoResearch (3 mentions)
Cx3cr1cre, by Jackson ImmunoResearch (2 mentions)
Dba 1j, by Jackson ImmunoResearch (2 mentions)
Ubc creert2, by Jackson ImmunoResearch (2 mentions)
Rag2 il2rg, by Taconic Biosciences (2 mentions)
Hif 1αfl fl, by Jackson ImmunoResearch (1 mentions)
Hif1αfl fl mice, by Jackson ImmunoResearch (1 mentions)
Bp0089, by BioXCell (1 mentions)
Cxcr2, by Jackson ImmunoResearch (1 mentions)
C57bl 6j wt mice, by Jackson ImmunoResearch (1 mentions)
Rosamt mg, by Jackson ImmunoResearch (1 mentions)
Cd11c cre, by Jackson ImmunoResearch (1 mentions)
10 protocols using mrp8 cre
1
Murine Caspase Knockout Models
2
Hypoxia-Inducible Factor-1α in Neutrophils
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Wild-type (WT) C57BL/6, CXCR2−/−, HIF-1αfl/fl and MRP8cre+ mice were obtained from The Jackson Laboratory, bred and maintained under specific pathogen-free (SPF) conditions in the Biological Resource Centre (BRC) of A*STAR, Singapore. HIF-1αfl/fl mice (The Jackson Laboratory, Bar Harbor, ME, USA) were a kind gift from Dr. Subhra K. Biswas at the Singapore Immunology Network. Male and female mice were used, and all experimental groups were gender-matched and age-matched. HIF-1αfl/fl and MRP8cre+ mice were crossed in-house to generate progeny with HIF-1α-deficient neutrophils (HIF-1αΔNφ). All experiments were performed under the approval of the Institutional Animal Care and Use Committee (IACUC, Singapore) of the BRC, in accordance with the guidelines of the Agri-Food and Veterinary Authority (AVA, Singapore) and the National Advisory Committee for Laboratory Animal Research (NACLAR, Singapore) of Singapore (IACUC protocol #171230). In some experiments, Ly-6G (Bio X Cell; Clone 1A8, BP0075) or isotype control antibody (Bio X Cell; BP0089) was administered via intraperitoneal (i.p.) injection at 200 ug per 20 g mouse three times per week.
3
Conditional Elmo1 Knockout Mice
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C57BL/6J, DBA/1J, NOD, Mrp8-Cre, Cx3cr1-Cre, Lyz2-Cre, and Ubc-CreERt2 mice were obtained from Jackson Laboratories. Elmo1fl/fl and Elmo1–/– mice have been described previously24 (link). To generate mice with deletion of Elmo1 in myeloid cells, neutrophils or macrophages, Elmo1fl/fl mice were crossed to Lyz2-Cre, Mrp8-Cre or Cx3cr1-Cre mice, respectively. To generate mice with inducible deletion of Elmo1, Elmo1fl/fl mice were crossed to Ubc-CreERt2. In these mice, Elmo1 deletion was induced by three daily intraperitoneal injections of 40 mg/kg tamoxifen dissolved in corn oil. To generate Elmo1–/–DBA mice, Elmo1–/– mice were backcrossed onto the DBA/1J background for at least 5 generations. KRN TCR transgenic mice22 (link) were a gift from Dr. Diane Mathis at the Harvard Medical School, and were bred to NOD mice to obtain the K/BxN mice23 (link), which develop progressive spontaneous arthritis. K/BxN serum was collected from 9-week old K/BxN mice by cardiac puncture. Age- and sex-matched littermate control animals were used for all experiments, and both males and females were assessed. In Elmo1fl/flLyz2-Cre mice, reduced disease severity was observed in female, but not in male mice (data not shown). All animal procedures were approved by and performed according to guidelines of the Institutional Animal Care and Use Committee (IACUC) at the University of Virginia.
4
Tracking Neutrophils in Murine Lungs
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Mice were housed and bred under specific pathogen–free conditions, and animal experiments were approved by the local authorities, conforming with ethical principles and animal protection laws at the respective location (reference 2.4.2.2 — 42/2018, State Agency Environment and Consumer Protection Saarland, Germany; IACUC protocol AN186887-01A, University of California, San Francisco, San Francisco, California, USA). Adam8−/− and Adam8+/+ mice were maintained on a C57BL/6J background for at least 10 generations, and C57BL/6J WT mice for ADAM8 inhibitor studies were purchased from Jackson Laboratory. To track neutrophils in the lung, MRP8Cre (stock 021614, Jackson Laboratory) mice were crossed with ROSAmT/mG (stock 007676, Jackson Laboratory) reporter mice. Weight-matched male and female mice aged 6–10 weeks were randomly allocated to experimental groups within a litter.
5
Genetic Manipulation of Mouse Inflammasome
6
Conditional Elmo1 Knockout Mice
7
Genetic Manipulation of Mouse Inflammasome
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Wild-type (WT) C57BL/6 (Jackson Laboratory), Casp1−/− (Rauch et al., 2017 (link)), Nlrc4−/−Asc−/− (Aachoui et al., 2015 (link)), Casp−/− (Kayagaki et al., 2011 (link)), Casp1−/−Casp11129mut/129mut referred to as Casp1−/−Casp11−/− (Kuida et al., 1995 (link)), Elane−/− (Jackson # 006112), Mpo−/− (Jackson Laboratory # 004265), Ncf1mt/mt referred to as Ncf1−/− (Jackson # 004742), Prf1−/− (Jackson # 002407), Rag1−/− (Jackson # 002216), Rag2−/−Il2rg−/− (Taconic # 4111), Gsdmd−/− (Rauch et al., 2017 (link)), Ifng−/− (Jackson # 002287), Casp11fl/fl (Cheng et al., 2017 (link)), Mrp8-cre (Jackson # 021614), and LysM-cre (Jackson # 004781) mice were used in this study. All mice were 6-12 weeks old, male or female, and housed under specific pathogen free condition-free facilities. All protocols were approved by the Institutional Animal Care and Use Committee at the University of Arkansas for medical Sciences at Little Rock, or Institutional Animal Care and Use Committee at the University of North Carolina at Chapel Hill.
8
Genetically Modified Mouse Models for Autophagy
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All flox mice (Atg5fl/lfl, Atg16l1fl/fl, and Becn1fl/fl) used in this study have been described previously [3 (link),87 (link)–89 (link)] and colonies are maintained in an enhanced barrier facility. LysM-Cre (Jax #004781), Cd11c-Cre (Jax #007567), Mrp8-Cre (Jax #021614) from the Jackson Laboratory were crossed to specific flox mice. Il17a-IRES-GFP-KI (Jax # 018472) reporter mice were bred to Atg5fl/fl-LysM-Cre and Atg5fl/fl mice to generate the Il17a-GFP/Atg5fl/fl-LysM-Cre and Il17a-GFP/Atg5fl/fl lines. Rubicon-/- (Jax # 032581) and WT littermates were provided by D. Green and J. Martinez [90 (link)]. Caspase 1/11-/- (Jax #016621) were bred to Atg5fl/fl-LysM-Cre and Becn1fl/fl-LysM-Cre mice. Parkin-/- (Jax # 006582) [91 (link)], Pink1-/- (Jax # 017946) [56 (link)], and WT control mice were a gift from Dr. Jonathan Brestoff at Washington University School of Medicine. Male and female littermates (aged 6 to 12 weeks) were used and were subject to randomization. A minimum of 3 mice were used per experiment and each experiment was performed twice. Statistical consideration was not used to determine mouse sample sizes. The mice were housed and bred at Washington University in St. Louis in specific pathogen-free conditions in accordance with federal and university guidelines, and protocols were approved by the Animal Studies Committee of Washington University.
9
Generating Myeloid-Specific LKB1 Knockout Mice
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Homozygous Stk11fl/fl mice (014143; Jackson Laboratory) [19 (link)] were crossed with LysM-cre [20 (link)] or Mrp8-cre mice (021614; Jackson Laboratory) [21 (link)] to generate mice that were deficient in myeloid-specific LKB1 (LysM-cre × Stk11fl/fl) and neutrophil-specific LKB1 (Mrp8-cre × Stk11fl/fl) [22 (link)]. Stk11fl/fl Cre-negative littermates were used as controls in all experiments. Experiments were approved by the Central Commission for Animal Experiments.
10
Genetic Manipulation of Lrp5 and Lrp6 in Mice
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All animal experiments were performed with the approval of Institutional Animal Care and Use Committee (IACUC) at Yale University. The LoxP-floxed Lrp5 (Lrp5fl/fl) and Lrp6 (Lrp6fl/fl) mice were obtained from Bart Williams57 (link). The Lrp5fl/fl and Lrp6fl/fl mice were back-crossed with C57BL/6 N mice for more than seven generations before being intercrossed with Lyz2-Cre (#004781), Rosa26-CreER2 (#008463), or Mrp8-Cre (#021614) mice (Jackson lab), followed by additional backcrossing with C57BL/6 N mice for more than ten generations. Ncr-1-Cre Lrp5fl/fl mice were described previously34 (link). Wildtype C57BL/6N mice were purchased from Envigo (#044). The mice were housed under specific-pathogen-free conditions in Yale Animal Resources Center facilities and under 12 h light/dark cycles at 68–79 °F and 30–70% humidity. Mice in all experiments were age- and gender-matched. Sample sizes used in animal experiments were based on the empirical determination in preliminary experiments. For deprivation of PUFAs, mice were placed on an essential fatty acid-free Diet (TD.84224, Envigo) for 4 days. For PUFA refeeding, 100 mg/kgbw PUFAs were delivered to mice four times via oral gavage every 12 h just before experiments.
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