fndc5 irisin  (Cusabio)

Journal: Npj Biological Physics and Mechanics

Article Title: Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice

doi: 10.1038/s44341-024-00006-y

Figure Lengend Snippet: a Representative images of immunohistochemistry stainings of total fibronectin using a polyclonal anti-fibronectin antibody (green), relaxed fibronectin fibers using Cy5.5-FnBPA5 (magenta) as well as cell nuclei using DAPI (blue) at all four timepoints (6 h p.i., 24 h p.i., 48 h p.i., and 72 h p.i.) and in the mock-infected mice (ctrl), demonstrate FnBPA5 signal in the muscularis externa at 24 h p.i. and in the mucosa at 72 h p.i. ( n = 5 mice per group). Scale bar: 100 µm. b Higher resolution zoom-ins of the same tissues for mucosa and muscularis externa highlights the kinetics of fibronectin fiber relaxation in the two cecal tissue layers. Scale bar: 50 µm. p.i., post infection; musc. ext., muscularis externa; L, lumen.

Article Snippet: Co-stainings for relaxed (Cy5.5-FnBPA5) and total fibronectin fibers as well as with anti-Ly6B.2 antibody (#MCA771G, BioRad, Hercules, California, USA) were performed as follows: Tissue cryosections (20 μm thick) on glass slides were encircled with a hydrophobic pen (H-4000, Vector Laboratories, USA) to decrease staining solution usage.

Techniques: Immunohistochemistry, Infection

Journal: Npj Biological Physics and Mechanics

Article Title: Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice

doi: 10.1038/s44341-024-00006-y

Figure Lengend Snippet: a Quantification of FnBPA5 + pixels in the mucosa yields a median rate of 8% of FnBPA5 + pixels at the late stage of the infection, while earlier days only show very low levels b: Quantification of FnBPA5 + pixels in the muscularis externa demonstrates a strong increase from the beginning of the inflammation, resulting in more than 80% of FnBPA5 + pixels at 72 h p.i. a + b FnBPA5 + pixels defined as pixels with intensity above the negative control value. Boxplots extend from first quartile to third quartile with median indicated as horizontal line and marker points representing individual mice ( n = 3–5). Statistical analysis: One-way ANOVA with Tukey’s multiple comparison test, p-values indicated. c + e Histograms of intensity distributions of FnBPA5 + pixels (c) (ctrl, 6 h, 24 h, 48 h, 72 h p.i. conditions from top to bottom) and FnBPA5/fibronectin intensity ratios (e) demonstrate that there is not only an increase in number of FnBPA5 + pixels but also a signal intensity increase of those in the late stage of the infection in the mucosa. d + f Histograms of intensity distributions of FnBPA5 + pixels (d) and FnBPA5/fibronectin intensity ratios (f) verify that there is not only an increase in number of FnBPA5 + pixels but also an increase in signal intensity. c–f Dashed vertical lines represent median intensity values. Pixel-by-pixel data pooled from at least 3 images from 3-5 mice per timepoint each. Fn, fibronectin.

Article Snippet: Co-stainings for relaxed (Cy5.5-FnBPA5) and total fibronectin fibers as well as with anti-Ly6B.2 antibody (#MCA771G, BioRad, Hercules, California, USA) were performed as follows: Tissue cryosections (20 μm thick) on glass slides were encircled with a hydrophobic pen (H-4000, Vector Laboratories, USA) to decrease staining solution usage.

Techniques: Infection, Negative Control, Marker, Comparison

Journal: Npj Biological Physics and Mechanics

Article Title: Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice

doi: 10.1038/s44341-024-00006-y

Figure Lengend Snippet: a Representative images of immunohistochemistry stainings of neutrophils using an anti-Ly6B.2 antibody (cyan), relaxed fibronectin fibers using Cy5.5-FnBPA5 (magenta) as well as cell nuclei using DAPI (blue) during the timecourse of the S . Tm infection (top to bottom: control mouse, 6 h, 24 h, 48 h and 72 h p.i.), show an increase of infiltrating neutrophil clusters in the mucosa from 24 h p.i. onwards but importantly the mucosal FnBPA5 signal is only increasing at 72 h p.i. (arrowheads). In contrast, the FnBPA5 signal in the muscularis mucosa and muscularis externa (arrows) is visible from 24 h p.i. (confirming results shown in Fig. ). Color-coded zoom-in overlay images for regions of interest are shown for every timepoint. Scale bar: 100 µm. Scale bar zoom-in: 50 µm. b Quantification of percentage of Ly6B.2 + cells in the mucosa during the timecourse demonstrates increasing numbers of neutrophils peaking at 48 h p.i. (25%) but keeping high (15%) levels of tissue neutrophils at 72 h p.i. c Quantification of FnBPA5 + pixels in spatial relation to neutrophil clusters reveals strong spatial correlation between them at 72 h p.i. At 48 h p.i., the percentage of FnBPA5 + pixels is comparable between mucosal areas with and without neutrophil clusters, while at 72 h p.i. the percentage of FnBPA5 + pixels in the neutrophil cluster regions is increased approximately 4-fold compared to areas without neutrophil clusters. b–c Marker points represent individual mice ( n = 3–5 mice per group with at least 10 images taken per mouse). Boxplots extend from first quartile to third quartile with median indicated as horizontal line. The whiskers represent the largest/lowest datapoint of the dataset that falls within 1.5× the interquartile range. Statistical analysis: One-way ANOVA with Tukey’s multiple comparison test, p-values indicated, n.s.: p ≥ 0.05. p.i., post infection.

Article Snippet: Co-stainings for relaxed (Cy5.5-FnBPA5) and total fibronectin fibers as well as with anti-Ly6B.2 antibody (#MCA771G, BioRad, Hercules, California, USA) were performed as follows: Tissue cryosections (20 μm thick) on glass slides were encircled with a hydrophobic pen (H-4000, Vector Laboratories, USA) to decrease staining solution usage.

Techniques: Immunohistochemistry, Infection, Control, Marker, Comparison

Journal: Npj Biological Physics and Mechanics

Article Title: Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice

doi: 10.1038/s44341-024-00006-y

Figure Lengend Snippet: a Schematic of LCM collection of FnBPA5-positive and FnBPA5-negative regions from the mucosa. Illustration generated using BioRender.com b Abundances of interesting differentially expressed proteins demonstrate significantly higher levels of aminopeptidase N (Apn) and tenascin C (Tnc) in the FnBPA5-positive regions. Statistical analysis: One sided student t-test, p-values corrected for multiple testing with Benjamin-Hochberg FDR correction, p values indicated, n.s.: p ≥ 0.05. Fn, fibronectin; Ngp, neutrophilic granule protein; Elane, neutrophil elastase.

Article Snippet: Co-stainings for relaxed (Cy5.5-FnBPA5) and total fibronectin fibers as well as with anti-Ly6B.2 antibody (#MCA771G, BioRad, Hercules, California, USA) were performed as follows: Tissue cryosections (20 μm thick) on glass slides were encircled with a hydrophobic pen (H-4000, Vector Laboratories, USA) to decrease staining solution usage.

Techniques: Generated

Journal: Npj Biological Physics and Mechanics

Article Title: Relaxation of mucosal fibronectin fibers in late gut inflammation following neutrophil infiltration in mice

doi: 10.1038/s44341-024-00006-y

Figure Lengend Snippet: a Schematic illustration of fibronectin's sequence with individual FnI, FnII and FnIII domains and binding regions indicated. b Schematic illustration with domain order and labeling adopted from UniProt: FnIII 8 corresponds to EDB, FnIII 13 corresponds to EDA. c: Sequence of peptides significantly enriched in the S . Tm infected cecum and their annotations on the fibronectin protein. d Relative abundances of the three identified peptides elevated in the inflamed cecum. Boxplots extend from first quartile to third quartile with marker points representing individual mice (2 mice per condition in technical duplicates) and horizontal lines the median abundance. The whiskers represent the largest/lowest datapoint of the dataset that falls within 1.5× the interquartile range. Statistical analysis: One sided student t-test, p-values corrected for multiple testing with Benjamin-Hochberg FDR correction, p-values indicated Fn, fibronectin.

Article Snippet: Co-stainings for relaxed (Cy5.5-FnBPA5) and total fibronectin fibers as well as with anti-Ly6B.2 antibody (#MCA771G, BioRad, Hercules, California, USA) were performed as follows: Tissue cryosections (20 μm thick) on glass slides were encircled with a hydrophobic pen (H-4000, Vector Laboratories, USA) to decrease staining solution usage.

Techniques: Sequencing, Binding Assay, Labeling, Infection, Marker