wt mice  (Charles River Laboratories)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: 1Platelet-driven leukotriene C4-mediated airway inflammation in mice is aspirin-sensitive and depends on T prostanoid receptors

doi: 10.4049/jimmunol.1402959

Figure Lengend Snippet: Effect of LTC 4 on eosinophilic pulmonary inflammation and platelet activation. OVA-sensitized C57BL/6 were challenged with 0.1% OVA on three successive days, with or without inhalation of LTC 4 (2.2 nmol) 12 h before each challenge. BAL fluid was collected

Article Snippet: WT C57BL/6 mice were purchased from Charles River.

Techniques: Activation Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: 1Platelet-driven leukotriene C4-mediated airway inflammation in mice is aspirin-sensitive and depends on T prostanoid receptors

doi: 10.4049/jimmunol.1402959

Figure Lengend Snippet: Effect of TP receptor deletion on LTC 4 -mediated eosinophil recruitment to the lung. A. Cytofluorographic detection of eosinophils and neutrophils (based on light scatter characteristics within the CD45+ gate) in the blood of representative C57BL/6 WT

Article Snippet: WT C57BL/6 mice were purchased from Charles River.

Techniques:

Journal: Molecular Metabolism

Article Title: Membrane metallo-endopeptidase (Neprilysin) regulates inflammatory response and insulin signaling in white preadipocytes

doi: 10.1016/j.molmet.2019.01.006

Figure Lengend Snippet: Subcutaneous and perigonadal MMEKO preadipocytes show impaired insulin signaling . MMEKO and control (wild-type C57BL/6) mice aged 12 weeks were sacrificed for in vitro insulin signaling. The stromal vascular cells (preadipocytes) from the subcutaneous (inguinal) and visceral (perigonadal) adipose depots were isolated. Cells were serum-starved for 4 h before a 20-minute insulin stimulation followed by protein isolation and western blot. (A) Western blot of protein of subcutaneous and perigonadal MMEKO preadipocytes showed impaired response among different insulin-signaling proteins. (B–I) Densitometric analysis of protein levels. The levels of unphosphorylated proteins were grouped by adipose depot (PG = Perigondal, SC = Subcutaneous) and genotype (black = Control, grey = MMEKO). The levels of phosphorylated proteins were grouped by adipose depot, genotype, and insulin stimulation (– = no insulin, + = 20 min after 100 μM insulin). Asterisks indicate p ≤ 0.05 by Student's t -test. N = 3. Bars indicate mean ± s.e.m.

Article Snippet: MMEKO mice were bred with WT C57BL/6 mice from Charles River Laboratories, and F2 homozygous male KO mice were used for subsequent experiments.

Techniques: Mouse Assay, In Vitro, Isolation, Western Blot

Journal: Nucleic Acids Research

Article Title: Oxidized dNTPs and the OGG1 and MUTYH DNA glycosylases combine to induce CAG/CTG repeat instability

doi: 10.1093/nar/gkw170

Figure Lengend Snippet: Steady-state levels of 8-oxodG and expression of the p53R2 protein in brain areas of wild-type and R6/2 mice. Groups of WT (n = 10) and R6/2 mice (n = 10) were sacrificed and levels of 8-oxo-dG were determined by HPLC-EC in genomic DNA prepared from the indicated brain areas. DNA 8-oxodG levels in ( A ) striatum and ( B ) motor cortex of 12 week-old WT (WT, open bar) and R6/2 mice (grey ba r ). Data are the mean±SE. The asterisks indicate significant differences by Student's t -test (* P

Article Snippet: Mice A colony of R6/2 ( ) transgenic and littermate wild-type (WT) mice was maintained at Charles River Laboratories (Calco, Italy).

Techniques: Expressing, Mouse Assay, High Performance Liquid Chromatography

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Invasive breast cancer reprograms early myeloid differentiation in the bone marrow to generate immunosuppressive neutrophils

doi: 10.1073/pnas.1424927112

Figure Lengend Snippet: Kinetics of G-CSF stimulation reveal G-CSF targets expansion of HSC and MPP populations in the BM. ( A and B ) BM, blood, spleen, lung, and MG (tumor) were harvested from WT FVB/n and PyMT mice (14–15 wk) or anti–G-CSF–treated mice. ( A ) Quantification of total BM cells ( Left ), cell numbers of Ly6G + and Ly6C hi cells ( Center ), and cell numbers of HSC, MPP F+ , and MPP F- populations ( Right ) in BM. ( B ) Representative images of femurs and tibias ( Left ) and spleens ( Right ). ( C–H ) BM, spleen, and blood were harvested from WT and G-CSF–stimulated (0.5–5 consecutive days) FVB/n mice. ( C ) Summary of the frequency (% total) of CD11b + Gr1 + cells assessed by flow cytometry. ( D ) FACS plots illustrate the frequency (% of total) of Ly6G + cells in BM, spleen, and blood ( Left ), histograms of Ly6G expression in Ly6G + cells ( Center ), and total cell numbers of Ly6G + cells ( Right ). ( E ) Cell numbers of CMPs, GMPs, and MEPs in BM. ( F ) Representative images of femurs and tibias. Cell numbers of ( G ) HSCs, MPPs F+ , and MPPs F- in BM and ( H ) HSCs, MPPs F+ , and MPPs F- in spleen. Data are representative of ( A and B ) three experiments (mean ± SEM, n = 3–7) and ( C–H ) two experiments (mean ± SEM, n = 3–10). * P

Article Snippet: WT FVB/n female mice were purchased from Charles River and WT C57BL/6 female (CD45.1 or CD45.2) mice were purchased from the Jackson (JAX) Laboratories for cross-breeding, in vivo G-CSF stimulation, or BM transplants.

Techniques: Mouse Assay, Flow Cytometry, Cytometry, FACS, Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Invasive breast cancer reprograms early myeloid differentiation in the bone marrow to generate immunosuppressive neutrophils

doi: 10.1073/pnas.1424927112

Figure Lengend Snippet: Breast cancer development results in profound remodeling of BM hematopoiesis. ( A ) Gating strategy to evaluate BrdU incorporation in Ly6G + cells by flow cytometry. ( B ) Representative FACS plots of BrdU incorporation (2 and 24 h after i.p. injection) in Ly6G + cells in BM and spleen of WT and PyMT FVB/n mice (14–15 wk). ( C ) Summary of the % of Ly6G + cells that are BrdU + in BM and spleen at 2 and 24 h. ( D ) Frequency (% of total) of CD11b + Gr1 + cells in BM of WT and PyMT FVB/n mice during tumor progression (8–15 wk). ( E ) Representative image of femurs and tibias from WT and PyMT FVB/n mice. ( F ) Schematic of myeloid differentiation. ( G–I ) The frequency and total cell numbers of MPs and HSPCs were evaluated in BM of WT and PyMT FVB/n mice (14–15 wk) by flow cytometry. ( G ) FACS plots illustrate the gating for MP, MPP, and HSC populations. Values shown are the frequency of cells within each parent gate. ( H ) Total cell numbers of MP subpopulations (CMP, GMP, and MEP). ( I ) Total cell numbers of HSCs and MPPs (MPP F- and MPP F+ ) in BM of WT and PyMT FVB/n mice (8–12 wk). Data are representative of ( A–C ) two experiments (mean ± SD, n = 3–4), ( D ) four experiments at each time point (mean ± SD, n = 10–12), ( E ) six experiments ( n = 8–15), ( H ) three experiments (mean ± SD, n = 5–7), ( I ) three experiments (8 and 10 wk; mean ± SD, n = 5), two experiments (12 wk; mean ± SD, n = 6–7), and five experiments (14–15 weeks; mean ± SD, n = 5–9). * P

Article Snippet: WT FVB/n female mice were purchased from Charles River and WT C57BL/6 female (CD45.1 or CD45.2) mice were purchased from the Jackson (JAX) Laboratories for cross-breeding, in vivo G-CSF stimulation, or BM transplants.

Techniques: BrdU Incorporation Assay, Flow Cytometry, Cytometry, FACS, Injection, Mouse Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Invasive breast cancer reprograms early myeloid differentiation in the bone marrow to generate immunosuppressive neutrophils

doi: 10.1073/pnas.1424927112

Figure Lengend Snippet: G-CSF is sufficient for the enhanced ROS activity and Rb1 low phenotype of tumor-induced Ly6G + neutrophils. ( A ) Superoxide production was assessed by DHR fluorescence in Ly6G + cells in blood from WT and isotype or anti–G-CSF–treated PyMT FVB/n mice (14–15 wk). Representative FACS plots illustrate the frequency (% of total) of Ly6G + cells ( Left ), histogram illustrates DHR fluorescence in Ly6G + cells ( Center ), and bar graphs summarize the MFI of DHR in Ly6G + cells ( Right ). Multiplex laser capture technology (Eve Technologies) was used to measure the concentration of G-CSF in the serum from ( B ) WT, PyMT, and anti–G-CSF–treated PyMT FVB/n mice (14–15 wk) and ( C ) PBS control treated or G-CSF–stimulated (12 or 24 h) WT mice. ( D ) Superoxide production was assessed by DHR fluorescence in Ly6G + cells in blood from WT, G-CSF–stimulated, and PyMT FVB/n mice (14–15 wk). Representative FACS plots illustrate the frequency (% of total) of Ly6G + cells ( Left ), histogram illustrates DHR fluorescence in Ly6G + cells ( Center ), and bar graphs summarize the MFI of DHR in Ly6G + cells ( Right ). ( E and F ) Rb1 expression was assessed by Western blot in ( E ) total splenocytes and ( F ) total splenocytes ( Upper ) or FACS-sorted Ly6G + cells from the spleen ( Lower ). WT or PyMT FVB/n mice (14–15 wk). ( G ) FACS plots illustrate the frequency (% of total) of Ly6G + cells and histrograms illustrate intracellular Rb1 expression in Ly6G + cells, assessed by flow cytometry. Data are representative of ( A ) two experiments (mean ± SEM, n = 3–4), ( B ) two experiments (mean ± SEM, n = 6), ( C ) one experiment ( n = 2–4), ( D ) two experiments (mean ± SEM, n = 3–4), ( E and F ) two experiments ( n = 2–3 biological samples), and ( G ) two experiments ( n = 4). * P

Article Snippet: WT FVB/n female mice were purchased from Charles River and WT C57BL/6 female (CD45.1 or CD45.2) mice were purchased from the Jackson (JAX) Laboratories for cross-breeding, in vivo G-CSF stimulation, or BM transplants.

Techniques: Activity Assay, Fluorescence, Mouse Assay, FACS, Multiplex Assay, Concentration Assay, Expressing, Western Blot, Flow Cytometry, Cytometry

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Invasive breast cancer reprograms early myeloid differentiation in the bone marrow to generate immunosuppressive neutrophils

doi: 10.1073/pnas.1424927112

Figure Lengend Snippet: Tumor-derived G-CSF, not GM-CSF or M-CSF, increases systemically during early tumor progression. Concentrations of G-CSF, GM-CSF, M-CSF, CXCL1, CCL2, CCL3, and CCL4 in ( A ) serum from WT and PyMT FVB/n mice (6–15 wk) and conditioned medium collected from ( B ) primary PyMT C57BL/6 tumor cells and ( C ) FVB/n VO-PyMT cell line were determined using multiplex laser bead technology (Eve Technologies). Data are representative of ( A ) two or three experiments (mean ± SEM, n = 4–7) and ( B and C ) two experiments, each sample in duplicate (mean ± SD of four biological samples). * P

Article Snippet: WT FVB/n female mice were purchased from Charles River and WT C57BL/6 female (CD45.1 or CD45.2) mice were purchased from the Jackson (JAX) Laboratories for cross-breeding, in vivo G-CSF stimulation, or BM transplants.

Techniques: Derivative Assay, Mouse Assay, Multiplex Assay