All mouse experiments were conducted in accordance with the Institutional Animal Care and Use Committee (IACUC) of Asan Institute for Life Sciences. Atg7fl/fl;LysM-Cre mice with myeloid cell-specific deletion of Atg7 were generated by crossing LysM-Cre mice (Jackson Laboratories, stock number 4781) with Atg7fl/fl mice (kindly provided by Prof. Masaaki Komatsu, Juntendo University, Tokyo, Japan). Atg7fl/fl littermate mice were used as control mice. Mice were genotyped with PCR using established primers [29 (link)].
Lysm cre mice
Manufactured by Jackson ImmunoResearch
Sourced in United States, Montenegro, Japan
LysM-Cre mice are a genetic model that expresses the Cre recombinase enzyme under the control of the lysozyme M (LysM) gene promoter. This allows for specific Cre-mediated recombination in myeloid cells, including macrophages and neutrophils.
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Lab products found in correlation
C57bl 6 mice, by Jackson ImmunoResearch (7 mentions)
Lysm cre, by Jackson ImmunoResearch (6 mentions)
C57bl 6j, by Jackson ImmunoResearch (6 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (6 mentions)
C57bl 6, by Jackson ImmunoResearch (4 mentions)
Trizol reagent, by Thermo Fisher Scientific (4 mentions)
C57bl 6 mice, by Charles River Laboratories (3 mentions)
Ccr2 mice, by Jackson ImmunoResearch (2 mentions)
C57bl 6j mice, by Charles River Laboratories (2 mentions)
Vav cre mice, by Jackson ImmunoResearch (2 mentions)
Rosa26 cas9 knockin mice, by Jackson ImmunoResearch (2 mentions)
B6.129 arntltm1bra j bmal1, by Jackson ImmunoResearch (2 mentions)
Clockδ19 mice, by Jackson ImmunoResearch (2 mentions)
C57bl 6 wt, by Jackson ImmunoResearch (2 mentions)
Tamoxifen, by Merck Group (2 mentions)
Ot 1 and ot 2 tcr transgenic mice, by Jackson ImmunoResearch (1 mentions)
Cd11c cre, by Jackson ImmunoResearch (1 mentions)
Batf3 mice, by Jackson ImmunoResearch (1 mentions)
Balb c, by Jackson ImmunoResearch (1 mentions)
Rapamycin, by Enzo Life Sciences (1 mentions)
105 protocols using lysm cre mice
1
Myeloid Cell-Specific Autophagy Regulation
2
Generation of ASXL1 Conditional Knockout Mice
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Asxl1‐MTfl/fl mice were generated as described previously17 and were crossed with Vav‐Cre mice,25 LysM‐Cre mice,26 and Lck‐Cre mice.27 LysM‐Cre mice were purchased from The Jackson Laboratory (stock no. 004781). Lck‐Cre mice were provided by the Laboratory Animal Resource Bank, National Institutes of Biomedical Innovation, Health, and Nutrition. MMTV‐PyMT mice (FVB/N background) were also purchased from The Jackson Laboratory (stock no. 002374) and backcrossed into the C57BL/6J (CLEA Japan) background for at least 12 generations.28 We generated Vav‐Cre; Asxl1‐MTfl/fl‐MMTV‐PyMT mice by crossing male MMTV‐PyMT mice with female Vav‐Cre; Asxl1‐MTfl/fl mice. Only female Vav‐Cre; Asxl1‐MTfl/fl‐MMTV‐PyMT or Asxl1‐MTfl/fl‐MMTV‐PyMT mice were used for experiments. All animal experiments were performed in accordance with the approved protocol (PA20‐26) from the Laboratory Animal Research Center of the Institute of Medical Science at the University of Tokyo.
3
Generation of ACE10/10-PPARα-LysM-Cre Mice
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To make ACE10/10-PPARα-floxed mice, WT PPARα-floxed mice [original made by Dr. Walter Wahli (18 (link)), University of Lausanne] were obtained from Dr. Mingyu Liang, Medical College of Wisconsin. These mice were crossed with ACE 10/10 mice. The A10-PPARα mice were further crossed with LysM-Cre mice (Jackson Labs strain #4781) (19 (link)) that was a gift of Dr. Gislaine Martins, Cedars-Sinai Medical Center. Eventually, we obtained ACE10/10-PPARα-LysM-Cre (A10-PPARα-Cre). The genetic design of the mice was represented in Figure 1A . Genotyping PCR was performed by using the primers shown in Supplemental Table 4. All mice were bred with 12 h day/night cycles and were allowed free access to food and water. Gender and age-matched adult mice (8–16 weeks) were used for each experiment. All animal experimental protocols were reviewed and approved by the Animal Welfare Committee Cedars-Sinai Medical Center (#8780).
4
Floxed Mouse Models for Immunology Studies
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Tsc1 floxed [51 (link)], mTor floxed [52 (link)], and Rptor floxed [53 (link)] mice; Cd11c-cre [54 (link)] and LysM-cre mice [55 (link)]; and OT-I and OT-II TCR transgenic mice [56 (link),57 (link)] were obtained from Jackson Laboratories. In all experiments, littermates carrying floxed alleles but without Cre recombinase were used as controls (WT).
5
Genetically Engineered Mouse Models
6
Generation and Characterization of Genetically Modified Mice
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BALB/c and C57BL/6 mice (female, 6–8 weeks old) were purchased from Jackson Laboratories (Bar Harbor, Maine, USA). A TFEBflox mouse line was generated using the ES cells from EUCOMM depository in Europe, which are the same cells used for generation of liver-specific TFEB knockout mice.24 (link) LysM-Cre mice were purchased from Jackson Laboratories and crossed with TFEBflox mice to generate MΦ-specific TFEB knockout (MΦ-TFEB-/-) mice. MΦ-specific TFEB transgenic (MΦ-TFEBtg) mice were generated and characterized previously.25 (link) Female FVB/N mice were bred with male heterozygous C3(1)/SV40Tag mice (a gift from Dr Jeffrey Green, National Cancer Institute). All mice were housed in the University of South Carolina or Washington University in St. Louis Animal Research Facilities.
7
Myeloid Cell-Specific Genetic Knockout
8
Genetic Murine Models in Tuberculosis Research
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TSC1f/f-ERCre+ mice were previously reported [27 (link), 56 (link)]. LysM-Cre+ mice were purchased from Jackson Laboratory. M. bovis (Bacillus Calmette-Guérin or BCG) was obtained from the laboratory of Dr. William Jacobs (Albert Einstein College of Medicine). Virulent M. tuberculosis H37Rv (ATCC, 25618D-2) was purchased from ATCC. Rapamycin was obtained from Enzo Life Science (Enzo Life Science, A275). Bafilomycin A1 (B1793), 3-methyladenine (3-MA, M9281), and wortmannin (W1628) were purchased from Sigma-Aldrich. Compound C (Millipore, 171,260) was obtained from Millipore (Billerica, MA).
9
Conditional Deletion of Pfkfb3 in Myeloid Cells
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Mice were used in accordance with the protocol approved by the Institutional Animal Care and Use Committee of Augusta University. The floxed Pfkfb3 (Pfkfb3flox/flox) mice were generated by Xenogen Biosciences Corporation (Cranbury, NJ, USA) (14 (link)). Myeloid-specific Pfkfb3 knockout was achieved by cross-breeding Pfkfb3flox/flox mice with Lysm-Cre mice (stock no. 004781, The Jackson Laboratory, Bar Harbor, ME, USA) to generate Pfkfb3flox/flox; Lysm-Cre (Pfkfb3ΔMϕ) mice. All mice were on a C57BL/6J background.
10
Conditional Knockout of Rheb1 in Mice
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C57BL/6 mice were used in this study. Lys-MCre mice were purchased from The Jackson Laboratories (Bar Harbor, ME, USA, stock NO. 004781). Rheb1flox/flox mice were kindly gifted by Professor Bo Xiao, Sichuan University. The mice were housed individually under standard conditions of temperature and humidity on a 12-h light/dark cycle. Lys-MCre mice were first mated with Rheb1flox/flox mice to generate Lys-MCre-Rheb1flox/+mice. Their offspring were then backcrossed to homozygote floxed mice (Lys-MCre-Rheb1flox/+X Rheb1flox/flox) to generate Lys-MCre-Rheb1flox/flox mice, hereafter, Lys-MCre-Rheb1flox/flox mice are referred to as homozygote KO mice and Rheb1flox/flox as wild-type (WT) mice. All animals were cared for according to the guidelines of the Southern Medical University Animal Care and Use Committee. All experimental protocols were approved by the Southern Medical University Animal Care and Use Committee.
We performed genotyping using genomic DNA isolated from mouse tail biopsies, and the primers are listed below:
Lys-MCre common: 5′-CTT GGG CTG CCA GAA TTT CTC3′; mutant: 5′-CCC AGA AAT GCC AGA TTA CG-3′; WT: 5′-TTA CAG TCG GCC AGG CTG AC-3′.
Rheb1-Forward: 5′-GCC CAG AAC ATC TGT TCC AT-3′; Rheb1-Reverse: 5′-GGT ACC CAC AAC CTG ACA CC-3′.
We performed genotyping using genomic DNA isolated from mouse tail biopsies, and the primers are listed below:
Lys-MCre common: 5′-CTT GGG CTG CCA GAA TTT CTC3′; mutant: 5′-CCC AGA AAT GCC AGA TTA CG-3′; WT: 5′-TTA CAG TCG GCC AGG CTG AC-3′.
Rheb1-Forward: 5′-GCC CAG AAC ATC TGT TCC AT-3′; Rheb1-Reverse: 5′-GGT ACC CAC AAC CTG ACA CC-3′.
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