Journal: Experimental Dermatology

Article Title: Itching Sensation and Elevated Interleukin‐31 Levels as Potential Indicators of Exceptional Response to Biologics in Patients With Moderate‐To‐Severe Psoriasis

doi: 10.1111/exd.70138

Figure Lengend Snippet: Correlation between serum (a) IL‐31, (b) LCN2 and (c) CCL2 levels and itching intensity in psoriasis patients. Each dot represents an individual patient sample ( n = 20). * p < 0.05; *** p < 0.001. CCL2, chemokine ligand 2; IL‐31, interleukin‐31; LCN2, lipocalin‐2; ns, not significant.

Article Snippet: ELISA kits for human IL‐31, LCN2 and CCL2 were purchased from ABclonal (Human Interleukin 31 ELISA kit, RK00378; Woburn, MA, USA), Origene Technologies (human NGAL ELISA kit, EA200004; Rockville, MD, USA) and R&D Systems (Human CCL2/MCP‐1 Quantikine ELISA kit, DCP00; Minneapolis, MN, USA).

Techniques:

Journal: Experimental Dermatology

Article Title: Itching Sensation and Elevated Interleukin‐31 Levels as Potential Indicators of Exceptional Response to Biologics in Patients With Moderate‐To‐Severe Psoriasis

doi: 10.1111/exd.70138

Figure Lengend Snippet: Differential serum pruritus‐associated protein levels in ER and non‐ER patients with psoriasis before and after treatment. Serum (a) IL‐31, (b) LCN2 and (c) CCL2 concentrations in the ER and non‐ER groups before and after treatment. Changes in (d) IL‐31, (e) LCN2 and (f) CCL2 concentrations according to biologics treatment in ER and non‐ER patients. Each dot represents an individual patient sample, n = 10 for each group. * p < 0.05; ** p < 0.01. CCL2, chemokine ligand 2; ER, exceptional responder; IL‐31, interleukin‐31; LCN2, lipocalin‐2; non‐ER, non‐exceptional responder; Tx, Treatment.

Article Snippet: ELISA kits for human IL‐31, LCN2 and CCL2 were purchased from ABclonal (Human Interleukin 31 ELISA kit, RK00378; Woburn, MA, USA), Origene Technologies (human NGAL ELISA kit, EA200004; Rockville, MD, USA) and R&D Systems (Human CCL2/MCP‐1 Quantikine ELISA kit, DCP00; Minneapolis, MN, USA).

Techniques:

Journal: Biological & pharmaceutical bulletin

Article Title: Piperacillin Exacerbates Vancomycin-Induced Toxicity in Renal Proximal Tubular Cells.

doi: 10.1248/bpb.b24-00726

Figure Lengend Snippet: Fig. 5. Changes in the mRNA Expression of Various AKI Biomarkers after VCM Treatment (A–D), and Concurrent Treatment with PIPC Enhanced the VCM-Induced Increase in (E) mRNA Expression and (F) NGAL Secretion in HK-2 Cells

Article Snippet: NGAL Enzyme-Linked Immunosorbent Assay NGAL secretion was determined using a Proteintech® Human NGAL enzyme-linked immunosorbent assay (ELISA) kit (Proteintech, Rosemont, IL, U.S.A.).

Techniques: Expressing

Journal: Heliyon

Article Title: Aloe-emodin alleviates inflammatory bowel disease in mice by modulating intestinal microbiome homeostasis via the IL-4/IL-13 axis

doi: 10.1016/j.heliyon.2024.e34932

Figure Lengend Snippet: AE alleviates mice colitis induced by DSS. A : Changes in body weights of mice were monitored following treatment with different doses of AE in a mouse model of colitis induced by DSS. B : Intestinal inflammation marker LCN2 in feces. Levels of LCN2 in fecal samples collected from mice were measured as a marker of intestinal inflammation. C : H&E staining of colon tissue sections. Histological evaluation of colon tissue slices stained with H&E to assess tissue damage and inflammation. Representative images are shown. Data are shown as mean ± SD, ***P < 0.001: compared with control group; ##P < 0.01 and ###P < 0.001: compared with DSS group. Representative results from one out of three independent experiments.

Article Snippet: Additionally, fecal samples collected on day 15 were used for the detection of lipocalin-2 (LCN2) using an ELISA kit (RK-KOA0515; Rockland, Limerick, PA, USA).

Techniques: Marker, Staining, Control

Journal: Heliyon

Article Title: Aloe-emodin alleviates inflammatory bowel disease in mice by modulating intestinal microbiome homeostasis via the IL-4/IL-13 axis

doi: 10.1016/j.heliyon.2024.e34932

Figure Lengend Snippet: The content of LCN2 in control, DSS induced, and AE treated mice (ng/gfecal).

Article Snippet: Additionally, fecal samples collected on day 15 were used for the detection of lipocalin-2 (LCN2) using an ELISA kit (RK-KOA0515; Rockland, Limerick, PA, USA).

Techniques: Control

Journal: Cell reports

Article Title: Lipocalin-2 regulates astrocyte-oligodendrocyte interaction to drive post-stroke secondary demyelination.

doi: 10.1016/j.celrep.2025.115899

Figure Lengend Snippet: Figure 2. Contralateral LCN2 elevation arises from local astrocytes and trans-callosal spread (A and B) Representative images and quantifications showing time-course changes of astrocytic LCN2 in the contralateral corpus callosum (n = 5 mice; scale bar, 20 μm). (C and D) Immunoblotting and quantitative analysis of LCN2 (n = 5 mice).

Article Snippet: Serum LCN2 was evaluated according to the manufacturer’s directions of LCN2 ELISA kit (FEK0853, Boster Biological Technology, China).

Techniques: Western Blot

Journal: Cell reports

Article Title: Lipocalin-2 regulates astrocyte-oligodendrocyte interaction to drive post-stroke secondary demyelination.

doi: 10.1016/j.celrep.2025.115899

Figure Lengend Snippet: Figure 4. Extracellular LCN2 gradually accumulates within mature oligodendrocytes (A and B) Representative images and quantifications showing LCN2 (red) surrounded by and accumulated into mature oligodendrocytes (GST-pi + , green; Olig2 + , blue) in the contralateral corpus callosum after dMCAO (n = 5 mice; scale bar, 20 μm). (C) Experimental flowchart in vivo. (D and E) Representative images and quantitative analysis of GFP (green) reporter LV into astrocytes (GFAP, blue) with induced hLCN2 (red) in the corpus callosum of Lcn2 − /− mice. White arrows indicate transfected astrocytes (scale bar, 20 μm).

Article Snippet: Serum LCN2 was evaluated according to the manufacturer’s directions of LCN2 ELISA kit (FEK0853, Boster Biological Technology, China).

Techniques: In Vivo, Transfection

Journal: Cell reports

Article Title: Lipocalin-2 regulates astrocyte-oligodendrocyte interaction to drive post-stroke secondary demyelination.

doi: 10.1016/j.celrep.2025.115899

Figure Lengend Snippet: Figure 7. Oligodendrocyte Lrp2 knockdown suppresses LCN2-related white matter degeneration (A) Relative mRNA expression levels of LCN2 receptors (Lrp2, 24p3r, Lrp6, Mc1r, Mc3r, Mc4r) (n = 5 independent primary cell cultures). (B and C) Representative fluorescent images and quantification of LRP2 expression in cultured oligodendrocytes (n = 5 mice; scale bar, 20 μm).

Article Snippet: Serum LCN2 was evaluated according to the manufacturer’s directions of LCN2 ELISA kit (FEK0853, Boster Biological Technology, China).

Techniques: Knockdown, Expressing, Cell Culture

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

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Neutrophil Gelatinase–Associated Lipocalin Protects from ANCA-Induced GN by Inhibiting T H 17 Immunity

doi: 10.1681/ASN.2019090879

Figure Lengend Snippet: NGAL is increased in mice with ANCA-induced GN and is released by ANCA-activated murine and human neutrophils. (A) Circulating and urinary NGAL levels detected by ELISA were increased in mice with anti-MPO–induced NCGN (AAV) compared with control mice (Ctrl). (B) Representative immunoblots show strong NGAL protein expression in murine neutrophils (Neutro), but not in monocytes (Mono) that were isolated by cell sorting. Actin served as loading control. (C) NGAL protein by ELISA is increased in supernatants of murine neutrophils primed with TNFα and stimulated with 125 µg/ml anti-MPO IgG. Stimulation with murine anti-BSA IgG and 25 ng/ml PMA, respectively, served as controls. (D) Representative immunoblots show strong NGAL protein expression in human neutrophils, but not in monocytes that were purified by cell sorting. Actin served as loading control. (E) TNFα-primed human neutrophils were stimulated with three different human MPO-ANCA, PR3-ANCA, or control IgG preparations (125 µg/ml), respectively. Treatment with either 10−6 fMLP or buffer (Buf) served as controls. ANCA caused a strong release of neutrophil-specific NGAL 46-kDa homodimers into the supernatant by immunoblotting. A representative immunoblotting experiment is depicted together with the corresponding statistics of all experiments. (F) NGAL by ELISA is increased in supernatants of ANCA-stimulated human neutrophils treated as in (E). ***P>0.001.

Article Snippet: Mouse IL-17 and mouse IFN γ Quantikine ELISA Kits (R&D Systems) were used to quantify IL-17A and IFN γ levels in supernatants from splenocytes stimulated with phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (1 μ g/ml) for 4 hours in the presence of 1× Brefeldin A. NGAL concentration in murine and human serum and urine as well as in supernatants from stimulated neutrophils was determined with the NGAL (LCN2) ELISA Kit from Dianova (Hamburg, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Isolation, FACS, Purification

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Neutrophil Gelatinase–Associated Lipocalin Protects from ANCA-Induced GN by Inhibiting T H 17 Immunity

doi: 10.1681/ASN.2019090879

Figure Lengend Snippet: NGAL−/− chimeric mice develop more severe anti-MPO–induced NCGN, but show similar NGAL levels in serum and urine compared with WT mice. (A) Experimental scheme depicting the induction of NCGN in MPO-deficient (MPO−/−) mice immunized with murine MPO (mMPO) that were subsequently irradiated and transplanted with either WT or NGAL-deficient (NGAL−/−) BM cells. All chimeric mice were analyzed 7–8 weeks after transplantation. (B) NGAL−/− chimeric mice developed more severe NCGN than WT chimeric mice with (C) higher percentage of crescents and necrosis. Shown are representative images of kidney sections stained with Periodic acid–Schiff at low (20×) and high (40×) magnification and quantitative analyses of glomeruli with crescents and necrosis. (D) Urine analysis reveals more erythrocyturia (Ery) and proteinuria (Prot) in NGAL−/− chimeric mice by dipstick and (E) higher albuminuria by ELISA, whereas (F) serum and urinary NGAL levels were similar in WT and NGAL−/− chimeric mice by ELISA. (G) Anti-MPO titer determined by ELISA was similar in WT and NGAL−/− chimeric mice. Ctrl, control; Leuk, leukocyturia. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Mouse IL-17 and mouse IFN γ Quantikine ELISA Kits (R&D Systems) were used to quantify IL-17A and IFN γ levels in supernatants from splenocytes stimulated with phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (1 μ g/ml) for 4 hours in the presence of 1× Brefeldin A. NGAL concentration in murine and human serum and urine as well as in supernatants from stimulated neutrophils was determined with the NGAL (LCN2) ELISA Kit from Dianova (Hamburg, Germany).

Techniques: Irradiation, Transplantation Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Neutrophil Gelatinase–Associated Lipocalin Protects from ANCA-Induced GN by Inhibiting T H 17 Immunity

doi: 10.1681/ASN.2019090879

Figure Lengend Snippet: NGAL−/− chimeric mice show similar renal myeloid cell, but stronger TH17 cell influx and increased renal CCL20 expression. MPO−/− mice immunized with murine MPO were irradiated and transplanted with either WT or NGAL−/− BM cells as described in Figure 2A. All chimeric mice were analyzed 7–8 weeks after transplantation. (A) Representative FACS plot with gating strategy and corresponding quantification of renal CD45+CD11b+Ly6G+ neutrophils (neutro) and CD45+CD11b+Ly6Chi classic monocytes (CM). (B) Representative FACS plots showing IL-17A and IFNγ expression by renal CD4+ T cells after PMA/ionomycin stimulation ex vivo. Quantification reveals increased renal total CD4+, CD4+ TH1 (IL-17A−IFNγ+) and CD4+ TH17 (IL-17A+IFNγ−) T cells in NGAL−/− compared with WT chimeric mice. (C) Immunohistochemistry of kidney sections stained for the T cell marker CD3 shows that most of the CD3+ T cells were detected within and around affected glomeruli (bold arrow), but not in nonaffected glomeruli (dotted arrow). (D) Quantification of renal CCL20 and IL1β mRNA by quantitative PCR with HPRT used as reference gene and renal CCL20 and IL1β protein by ELISA. Both renal CCL20 mRNA and protein were increased in mice transplanted with NGAL−/− BM. (E) Splenic cells from WT and NGAL−/− chimeric mice were stimulated with PMA/ionomycin ex vivo and IFNγ and IL-17A was determined in the supernatants by ELISA. Cells isolated from mice transplanted with NGAL−/− BM produced more IFNγ and IL-17A compared with WT BM-transplanted mice. *P<0.05; **P<0.01.

Article Snippet: Mouse IL-17 and mouse IFN γ Quantikine ELISA Kits (R&D Systems) were used to quantify IL-17A and IFN γ levels in supernatants from splenocytes stimulated with phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (1 μ g/ml) for 4 hours in the presence of 1× Brefeldin A. NGAL concentration in murine and human serum and urine as well as in supernatants from stimulated neutrophils was determined with the NGAL (LCN2) ELISA Kit from Dianova (Hamburg, Germany).

Techniques: Expressing, Irradiation, Transplantation Assay, Ex Vivo, Immunohistochemistry, Staining, Marker, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Isolation, Produced

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Neutrophil Gelatinase–Associated Lipocalin Protects from ANCA-Induced GN by Inhibiting T H 17 Immunity

doi: 10.1681/ASN.2019090879

Figure Lengend Snippet: IL-17A gene deficiency rescues NGAL−/− chimeric mice from aggravated NCGN. (A) Experimental scheme for the induction of anti-MPO induced NCGN in WT, NGAL−/−, and IL-17A−/−/MPO−/− chimeric mice. MPO-immunized MPO−/− mice were irradiated and transplanted with either WT or NGAL-deficient (NGAL−/−) BM. Murine MPO-immunized IL-17A−/−/MPO−/− mice were irradiated and transplanted with NGAL−/−/IL-17A−/− BM. All chimeric mice were analyzed 7–8 weeks after transplantation. (B) Representative image of kidney sections stained with Periodic acid–Schiff at low (20×) and high (40×) magnification. Glomeruli with crescents and necrosis were increased in NGAL−/− chimeric mice compared with WT chimeric mice and reduced in the NGAL−/−/IL-17A−/− chimeric mice. (C) Leukocyturia (Leuk), proteinuria (Prot), and erythrocyturia (Ery) were measured in urine by dipstick. (D) Albuminuria was quantified by ELISA. (E) Serum and urinary NGAL levels were measured by ELISA. (F) Renal influx of neutrophils, classic monocytes (CM), and CD4+ T cells was increased in NGAL−/− chimeric mice compared with WT mice and reduced in the NGAL−/−/IL-17A−/− mice. Immune cells were identified using specific markers by flow cytometry. Total number of neutrophils, CM, and CD4+ T cells per kidney are shown. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Mouse IL-17 and mouse IFN γ Quantikine ELISA Kits (R&D Systems) were used to quantify IL-17A and IFN γ levels in supernatants from splenocytes stimulated with phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (1 μ g/ml) for 4 hours in the presence of 1× Brefeldin A. NGAL concentration in murine and human serum and urine as well as in supernatants from stimulated neutrophils was determined with the NGAL (LCN2) ELISA Kit from Dianova (Hamburg, Germany).

Techniques: Irradiation, Transplantation Assay, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry