Journal: Mucosal immunology

Article Title: β-Glucans in food modify colonic microflora by inducing antimicrobial protein, calprotectin, in a Dectin-1-induced-IL-17F-dependent manner.

doi: 10.1038/mi.2017.86

Figure Lengend Snippet: Figure 3 Interleukin-17F (IL-17F) production is decreased in colonic lamina propria (cLP) of Clec7a / mice. (a) Cytokine gene expression in cLP was examined by quantitative real-time reverse transcription PCR (qRT-PCR), and expression levels in Clec7a / mice are indicated relative to those in wild-type (WT) mice (n ¼ 3–8/group). (b,c) IL-17F þCD45 þ cell population in cLP layer from WT or Clec7a / mice was examined by flow cytometry. (n ¼ 4–5/group). (d) S100A8 þ and IL-17F þ cells in whole colonic cells from WT mice were examined by flow cytometry. Data are representatives of two independent experiments, and in a and c are expressed as mean±s.d. *Po0.05 vs. WT control.

Article Snippet: Antibodies against mouse CD11b (M1/70), CD11c (HL3), CD45 (30-F11), Ly6G (1A8), and Ly6C (HK1.4) were obtained from BioLegend (San Diego, CA), antibodies against IL-17F (18F10) and EpCAM-1 (G8.8) were obtained from eBioscience (San Diego, CA), and antibody against S100A8 (cat. no. BAF3059) was obtained from R&D Systems.

Techniques: Gene Expression, Reverse Transcription, Quantitative RT-PCR, Expressing, Flow Cytometry, Control

Journal: Mucosal immunology

Article Title: β-Glucans in food modify colonic microflora by inducing antimicrobial protein, calprotectin, in a Dectin-1-induced-IL-17F-dependent manner.

doi: 10.1038/mi.2017.86

Figure Lengend Snippet: Figure 6 Dectin-1 ligands are not detected in intestinal commensal microbiota. (a) Wild-type (WT) mouse fecal microbiota or Candida albicans (SC5314) were pretreated with non-immune goat or rabbit serum to block nonspecific binding. Then, they were treated with PE-labeled b-glucan- recognizing protein (Pi-BGRP) or the mutant form of the b-glucan recognition site of Pi-BGRP (Pi-BGRP.M), and PE þ population was determined by flow cytometry. (b) Thioglycollate-induced peritoneal macrophages (Mfs) from WT, Clec7a / , Myd88 / , or Clec7a / Myd88 / mice were cocultured with live Alcaligenes faecalis (multiplicity of infection (MOI): 1 or 10) for 24 h and the tumor necrosis factor-a (TNF-a) concentration in the culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA; n ¼ 3/group). (c) S100A8 expression in colonic epithelial cells (cECs) from A. faecalis monocolonized (A. faecalis), germ-free (GF), or specific-pathogen-free SPF mice was examined by flow cytometry (n ¼ 3–4/group). Data in b and c are representatives of two independent experiments and are expressed as mean±s.d.

Article Snippet: Antibodies against mouse CD11b (M1/70), CD11c (HL3), CD45 (30-F11), Ly6G (1A8), and Ly6C (HK1.4) were obtained from BioLegend (San Diego, CA), antibodies against IL-17F (18F10) and EpCAM-1 (G8.8) were obtained from eBioscience (San Diego, CA), and antibody against S100A8 (cat. no. BAF3059) was obtained from R&D Systems.

Techniques: Blocking Assay, Binding Assay, Labeling, Mutagenesis, Flow Cytometry, Infection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Biomolecules

Article Title: Evaluation of Plasma-Derived hsa_circ_003077 for Non-Invasive Diagnosis of Alzheimer’s Disease

doi: 10.3390/biom16030356

Figure Lengend Snippet: Plasma glial and receptor biomarkers in Alzheimer’s disease (AD) patients and cognitively healthy controls (HC). ( a ) GFAP, ( b ) NfL, ( c ) Tyro3, and ( d ) AXL levels were measured in plasma. Data are presented as mean ± SEM. Statistical comparisons were performed using the Mann–Whitney U test. Statistical significance is indicated as ** p < 0.01; *** p < 0.001; ns, not significant.

Article Snippet: The Human Tyro3 DuoSet ELISA kit (Catalog No: DY8596-05) supplied by R&D Systems (Minneapolis, MN, USA) was used to measure Tyro3 levels, while the Human AXL DuoSet ELISA kit (Catalog No: DAXL00; R&D Systems, Minneapolis, MN, USA) was used to measure AXL levels.

Techniques: Clinical Proteomics, MANN-WHITNEY

Journal: Biomolecules

Article Title: Evaluation of Plasma-Derived hsa_circ_003077 for Non-Invasive Diagnosis of Alzheimer’s Disease

doi: 10.3390/biom16030356

Figure Lengend Snippet: Diagnostic performance of plasma biomarkers for differentiating Alzheimer’s disease (AD) from cognitively healthy controls (HC) based on ROC curve analysis. Receiver operating characteristic (ROC) curves were generated to assess the diagnostic accuracy of plasma biomarkers. The area under the ROC curve (AUC) was used to evaluate biomarker performance: 0.90–1.00, superior diagnostic efficacy; 0.80–0.89, good diagnostic efficacy; 0.70–0.79, moderate diagnostic efficacy; <0.70, poor/substandard diagnostic efficacy. Data are presented as mean ± SEM. Statistical comparisons were performed using the Mann–Whitney U test. hsa_circ_003077 exhibited the highest diagnostic accuracy (AUC = 0.90; 95% CI: 0.82–0.97), followed by NfL (AUC = 0.75) and pTau-217 (AUC = 0.77). AXL and GFAP showed lower diagnostic performance (AUC = 0.63 and 0.69, respectively), while pTau-181 and Tyro3 demonstrated substandard efficacy (AUC = 0.63 each).

Article Snippet: The Human Tyro3 DuoSet ELISA kit (Catalog No: DY8596-05) supplied by R&D Systems (Minneapolis, MN, USA) was used to measure Tyro3 levels, while the Human AXL DuoSet ELISA kit (Catalog No: DAXL00; R&D Systems, Minneapolis, MN, USA) was used to measure AXL levels.

Techniques: Diagnostic Assay, Clinical Proteomics, Generated, Biomarker Discovery, MANN-WHITNEY

Journal: Inflammation research : official journal of the European Histamine Research Society ... [et al.]

Article Title: A positive feedback cycle between the alarmin S100A8/A9 and NLRP3 inflammasome-GSDMD signalling reinforces the innate immune response in Candida albicans keratitis.

doi: 10.1007/s00011-023-01757-5

Figure Lengend Snippet: Fig. 6 Alarmin S100A8/A9 forms a positive feedback loop with NLRP3 inflammasome- GSDMD in the pathogenesis of Candida albicans keratitis

Article Snippet: Primary antibodies targeting S100A8 (1:250; #MAB3059, R&D systems), S100A9 (1:500; #ab242945, Abcam), TLR4 (1:500; #ab13556, Abcam), NLRP3 (1:500; #ab263899, Abcam), cleaved CASP1 (1:250; #89,332, Cell Signaling Technology), ASC (1:250; #sc-514414, Santa Cruz Biotechnology), GSDMDC1 (1:250; #sc-393581, Santa Cruz Biotechnology), and β-actin (1:1000; #AC026, ABclonal) were used in this study.

Techniques:

Journal: Immunity

Article Title: Tumor necrosis factor-α blocks differentiation and enhances suppressive activity of immature myeloid cells during chronic inflammation.

doi: 10.1016/j.immuni.2013.02.007

Figure Lengend Snippet: Figure 5. TNF-a Blocks MDSC Maturation via S100A8 and S100A9 and RAGE Proteins (A) S100A8 and S100A9 mRNA expression in splenocytes are shown relative to Tnf+/+ noninflamed mice, set as 1. (B) S100A8 and S100A9 mRNA expression in isolated spleen-derived MDSCs is shown relative to Tnf+/+ MDSCs (set as 1). Data of two independent experiments are presented (mean ± SEM). (C) S100A9, S100A8, and RAGE protein expression in splenic MDSCs isolated from inflamed mice were evaluated by immunoblotting. a-Tubulin served as a control. (D) S100A9, S100A8, and RAGE expression in splenic MDSCs isolated from inflamed Tnf/ mice incubated with GM-CSF and TNF-a for 3 days were evaluated by immunoblotting. a-Tubulin served as a control. (E) MDSCs isolated from spleens of inflamed Tnf/ mice were cultured with GM-CSF, TNF-a, RAGE antibody, and S100A8 and S100A9 antibodies (together or separately) for 5 days, and cells’ phenotype was evaluated. Results are representative of two or more independent experiments (error bars show SD, n = 4–9 per group). *p < 0.01; **p < 0.002 (t test). See also Figure S5.

Article Snippet: In some experiments, 100 ng/ml of TNF-a (PeproTech) and a mixture of 10 mg/ml of RAGE (AF1179; R&D Systems) and 20 mg/ml of S100A8 and S100A9 (AF3059 and AF2065; R&D Systems) neutralizing antibodies were added to the cells, which were incubated with GM-CSF.

Techniques: Expressing, Isolation, Derivative Assay, Western Blot, Control, Incubation, Cell Culture

Journal: PLoS ONE

Article Title: S100A8/A9 Is Not Involved in Host Defense against Murine Urinary Tract Infection

doi: 10.1371/journal.pone.0013394

Figure Lengend Snippet: S100A8/A9 protein expression in bladder (1A) and kidney (1B) homogenate from sham WT mice (white bar) and from WT mice 24 and 48 hours after UTI with 9×10 8 E. coli CFU/mouse (black bars). Data are mean ± SEM; N = 7–8 mice per group, *P<0.05, **P<0.005 versus sham, #P<0.005 versus 24 hours UTI. Sequential double AP staining using Ly6G (indicating granulocyte staining) and S100A9 antibodies on kidney tissue from sham WT mice (1C) or WT mice 24 hours after UTI with 9×10 8 E. coli CFU (1D–F). Using spectral imaging Ly6G/S100A9 double staining was separately visualized for Ly6G (Vector Blue, 1E) or S100A9 (Vector Red, 1F). Scale bars display 0.1 mm.

Article Snippet: Sections were incubated with rat-anti-mouse S100A8 or S100A9 (R&D systems, Abingdon, United Kingdom) for 3 hours followed by goat-anti-rat IgG AP.

Techniques: Expressing, Staining, Imaging, Double Staining, Plasmid Preparation

Journal: Molecular Vision

Article Title: Calcium-binding S100 protein expression in pterygium

doi:

Figure Lengend Snippet: Primary antibodies used in the experiment.

Article Snippet: S100A8 , Mouse monoclonal (8–5C2) , Acris Antibodies , 1/150 , 1/1000.

Techniques:

Journal: Molecular Vision

Article Title: Calcium-binding S100 protein expression in pterygium

doi:

Figure Lengend Snippet: Primers used in real-time PCR.

Article Snippet: S100A8 , Mouse monoclonal (8–5C2) , Acris Antibodies , 1/150 , 1/1000.

Techniques: Sequencing

Journal: Molecular Vision

Article Title: Calcium-binding S100 protein expression in pterygium

doi:

Figure Lengend Snippet: Immunofluorescent staining of S100 proteins in human conjunctival and pterygial epithelia. Primary antibodies against S100A4 ( C and D ), S100A6 ( E and F ), S100A8 ( G and H ), S100A9 ( I and J ), and S100A11 ( K and L ) were used. Negative controls were shown in A and B . Images of uninvolved conjunctiva ( A , C , E , G , I , and K ) and pterygium epithelium ( B , D , F , H , J , and L ) were shown. The nuclei were stained with DAPI present in the mounting medium. A dashed line indicates the location of the basement membrane. All images were taken at 400X magnification.

Article Snippet: S100A8 , Mouse monoclonal (8–5C2) , Acris Antibodies , 1/150 , 1/1000.

Techniques: Staining, Membrane

Journal: Molecular Vision

Article Title: Calcium-binding S100 protein expression in pterygium

doi:

Figure Lengend Snippet: Western blot analysis of S100 proteins expressed in human conjunctiva and pterygium tissue. Uninvolved conjunctiva (Lane 1) and pterygium (Lane 2) tissues were lysed in RIPA buffer and 60 μg of total protein was loaded in each lane of SDS-PAGE and transferred to nitrocellulose membrane and probed with antibodies against various S100 proteins as well as β-actin (loading control). Bands of 11 kDa corresponding to S100A4, S100A6, and S100A11 were detected. Bands corresponding to the molecular weight of 8 kDa and 13 kDa confirmed the presence of S100A8 and S100A9, respectively.

Article Snippet: S100A8 , Mouse monoclonal (8–5C2) , Acris Antibodies , 1/150 , 1/1000.

Techniques: Western Blot, SDS Page, Membrane, Control, Molecular Weight

Journal: Molecular Vision

Article Title: Calcium-binding S100 protein expression in pterygium

doi:

Figure Lengend Snippet: Fold difference of S100A4 , S100A6 , S100A8 , S100A9 , and S100A11 gene transcripts in pterygium tissue relative to conjunctiva tissue. Total RNA was extracted from uninvolved conjunctiva and pterygium tissue after mechanical homogenization. Reverse transcription for 0.5 μg of total RNA. Glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) was used as the internal control. ΔC t was calculated by subtracting the C t of GAPDH from the C t of the targeted gene. The fold change was determined by the ΔΔC t method (see text). The uninvolved conjunctiva sample was used as a reference for each of the tested S100 transcripts in the pterygium sample. Height of the bars represents mean value and error bars represent standard deviation. An asterisk indicates that p is less than 0.05 (p<0.05).

Article Snippet: S100A8 , Mouse monoclonal (8–5C2) , Acris Antibodies , 1/150 , 1/1000.

Techniques: Homogenization, Reverse Transcription, Control, Standard Deviation