Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Prenatal treatment with preimplantation factor improves early postnatal neurogenesis and cognitive impairments in a mouse model of Down syndrome

doi: 10.1007/s00018-024-05245-9

Figure Lengend Snippet: Treatment with sPIF restores DYRK1A and BDNF protein levels in the brain of Dp(16)1Yey pups on P6. Evaluation of the DYRK1A protein level in WT pups (vehicle: n = 5; sPIF: n = 5) and Dp(16)1Yey pups (vehicle: n = 3; sPIF: n = 5) ( A ). The brain BDNF protein concentration in WT pups (vehicle: n = 6; sPIF: n = 4) and Dp(16)1Yey pups (vehicle: n = 4; sPIF: n = 6) ( B ). Data are expressed as the mean ± SD and were analyzed in a two-way ANOVA followed by Fisher’s least squares difference test. ** p < 0.01

Article Snippet: The levels of brain-derived neurotrophic factor (BDNF) in brain lysates were measured with a mouse BDNF PicokineTM ELISA kit (Boster).

Techniques: Protein Concentration

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: Primer sequences for the real-time PCR amplification.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Real-time Polymerase Chain Reaction, Amplification

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: Elevated lipocalin 2 (Lcn2) during E. coli O157:H7 infection. (A) Real-time PCR analysis of the levels of Lcn2 mRNA expression in indicated tissues of mice. (B) The mRNA levels of Lcn2 expression in indicated tissues of control and E. coli O157:H7-infected mice. (C) Serum levels of Lcn2 protein concentration in E. coli O157:H7-infected mice after challenge at different time points. (D–G) The mRNA expression levels of Lcn2 in indicated tissues from E. coli O157:H7-infected mice after challenge at different time points. (H) Protein levels of Lcn2 in mice detected on immunohistochemistry sections of liver and jejunum at 8 and 32 h after challenge with E. coli O157:H7. Original magnification was 200×. Values are average means of triplicate experiments. Error bars depict SEM ( n = 4 per time point). Results are expressed as means ± SEM. * P < 0.05, and ** P < 0.01.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Infection, Real-time Polymerase Chain Reaction, Expressing, Control, Protein Concentration, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: The bacteriostatic characteristics of lipocalin 2 (Lcn2). (A,B) Bacterial loads in blood (CFU/ml) and livers (CFU/mg) of E. coli O157:H7-infected mice 32 hpi. (C,D) Serum levels of Lcn2 protein measured and growth of E. coli O157:H7 in RPMI with 20% acute-phase serum from wild-type (WT) or Lcn2 −/− mice. Values are average means of triplicate experiments with two mice per genotype per experiment. Error bars depict SEM ( n = 6 per group). Results are expressed as means ± SEM. P < 0.05 was considered statistically significant. * P < 0.05.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: Granulocyte abnormalities in Lcn2 −/− mice. (A) Hematological parameters of peripheral blood from wild-type (WT) and Lcn2 −/− mice. The data are presented as mean × 10 3 cells/μl. Error bars depict SEM ( n = 6 per group). (B) Flow cytometry analysis of neutrophils in the peripheral blood after intragastric administration with 2 × 10 8 CFU of E. coli O157:H7. Cells were stained with indicated clones of Gr-1 Ab, and positive cells were determined by flow cytometry. Values are average means of triplicate experiments with two mice per genotype per experiment. Error bars depict SEM. (C) Wright-Giemsa staining of peripheral blood smears from Lcn2 −/− mice identified atypical hyposegmented neutrophils in the peripheral blood. Original magnification ×63. In contrast, WT mice displayed normal neutrophil maturation. (D) Enumeration of the number of band neutrophils in the peripheral blood of Lcn2 −/− mice. The data are presented as mean band cell numbers per 100 leukocytes. Error bars depict SEM. Results are expressed as means ± SEM. P < 0.05 was considered statistically significant. * P < 0.05, and ** P < 0.01.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Flow Cytometry, Staining, Clone Assay

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: Reduced migration of lipocalin 2-deficient ( Lcn2 −/− ) neutrophils. (A,B) Flow cytometry analysis of peripheral blood and peritoneal exudates of heat-killed E. coli O157:H7-challenged mice following staining with a Gr-1 PE Ab. (C,D) ELISA analysis of TNF-α in the serum and peritoneal exudates of heat-killed E. coli O157:H7-challenged mice. (E,F) Quantitative determination of chemokines MCP-1 and MIP-2 mRNA expression in the liver of heat-killed E. coli O157:H7-challenged mice. Values are average means of triplicate experiments with two mice per genotype per experiment. Error bars depict SEM. Results are expressed as means ± SEM. P < 0.05 was considered statistically significant. * P < 0.05 and ** P < 0.01.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Migration, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: Decreased expression of inflammatory cytokines produced by lipocalin 2-deficient ( Lcn2 −/− ) macrophages. (A) Real-time PCR analysis of Lcn2 mRNA expression levels in the E. coli O157:H7-infected primary bone marrow-derived macrophages (BMDMs) from wild-type (WT) and Lcn2 −/− mice. (B–D) ELISA analysis of interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α levels in the culture medium of the E. coli O157:H7-infected BMDMs from WT and Lcn2 −/− mice. (E–L) Real-time PCR analysis of cytokine mRNA expression levels in the E. coli O157:H7-infected pBMDMs from WT and Lcn2 −/− mice. (M) The infected BMDMs of WT and Lcn2 −/− mice were subjected to staining with rabbit monoclonal antibody iNOS, rat monoclonal antibody F4/80, Alexa Fluor 488 goat anti-rabbit IgG, and Alexa Fluor 647 goat anti-rat IgG in blocking buffer (1:200) and observed by fluorescence microscopy. Values are average means of triplicate experiments with two mice used for the isolation of BMDMs per genotype per experiment. Error bars depict SEM. Results are expressed as means ± SEM. P < 0.05 was considered statistically significant. * P < 0.05 and ** P < 0.01.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Expressing, Produced, Real-time Polymerase Chain Reaction, Infection, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining, Blocking Assay, Fluorescence, Microscopy, Isolation

Journal: Frontiers in Immunology

Article Title: Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function

doi: 10.3389/fimmu.2019.02594

Figure Lengend Snippet: Lipocalin 2 (Lcn2) can promote migration and phagocytosis of macrophages. (A,B) Scratch wound healing assay of mouse RAW264.7 macrophages and quantification of the fold change of average migrated distance of cells was measured with microscope ( n = 3, mean ± SEM, scale bars, 100 μm). (C,D) ELISA analysis of monocyte chemoattractant protein (MCP)-1 and macrophage inflammatory protein (MIP)-2 levels in the culture medium of Lcn2-treated macrophages. (E) Flow cytometry analysis of mouse RAW264.7 macrophages incubated with FITC-dextran. Values are average means of triplicate experiments with two repetitions per treatment per experiment. Error bars depict SEM. * P < 0.05 and ** P < 0.01.

Article Snippet: Serum Lcn2 was quantified using Lcn2 Mouse ELISA kits (Boster, China) according to the manufacturer's instructions.

Techniques: Migration, Wound Healing Assay, Microscopy, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Incubation