recombinant mouse eotaxin  (R&D Systems)

Journal: bioRxiv

Article Title: Microbiome depletion rejuvenates the aging brain

doi: 10.64898/2026.02.13.705770

Figure Lengend Snippet: a, Representative immunofluorescence images of Iba1+ (magenta) and CD68+ (green) microglia in the cortex of aged mice treated with water or antibiotics (Abx); quantification shows a significant reduction in both Iba1 and CD68 immunoreactivity following microbiome depletion. Yellow arrows indicate double positive cells (n = 5/group). Scale bar, 50 μm. b, Schematic of the experimental design: aged mice were treated with Abx or water for 30 days, after which plasma and cortical lysates were collected for cytokine profiling using a proteome array. c, Heatmaps showing fold change in cytokine and chemokine levels in plasma (left) and cortical lysates (right) relative to water controls. Multiple inflammatory mediators were downregulated upon Abx treatment; Eotaxin-1 (CCL11), highlighted in red, was significantly reduced in both compartments. d, Violin plots showing fold change in Eotaxin-1 abundance in plasma (top) and brain (bottom), confirming a significant reduction in circulating and central levels following microbiome depletion (plasma, n = 6/group; brain, n = 3/group). e, Schematic of rescue experiment: aged mice received antibiotics (Abx) to deplete the microbiome, in combination with either vehicle or recombinant Eotaxin-1 (n = 5/group). f-h, Quantification of blood vessel density (g), BrdU+ cells in the dentate gyrus (h), and CD68+ area in the cortex (i) (n = 5/group). All data represented as mean ± s.e.m.; * P < 0.05, ** P < 0.01, **** P < 0.0001, Student’s t -test (a,d); ANOVA with Tukey’s multiple-comparisons post hoc test (f-h).

Article Snippet: For overexpression experiments, a separate cohort of 23-month-old C57BL/6J mice was treated with antibiotics for 30 days (as above) and concurrently injected intraperitoneally with recombinant mouse Eotaxin (10 ng/g body weight in PBS (2 μg/ml stock; R&D Systems, 420ME020) every 3 days.

Techniques: Immunofluorescence, Clinical Proteomics, Recombinant

Journal: bioRxiv

Article Title: Microbiome depletion rejuvenates the aging brain

doi: 10.64898/2026.02.13.705770

Figure Lengend Snippet: a, Schematic of the experimental design: aged mice were treated with either control IgG or an anti–eotaxin-1 neutralizing antibody for 30 days (Control, n = 3, Eotaxin Ab, n = 5). b, Representative CD31 immunostaining and quantification showing increased blood vessel density in the cortex of Eotaxin-1–inhibited mice. c, BrdU labeling (green, arrows) of proliferating cells in the DG and quantification of BrdU+ cells reveals enhanced adult neurogenesis following Eotaxin-1 inhibition. d, Immunofluorescence for Iba1 (magenta) and CD68 (green) shows reduced microglial activation in Eotaxin-1–treated mice. Yellow arrows indicate double positive cells. e, Quantification of myelinating oligodendrocyte precursor cells (Pdgfra+, green) in the corpus callosum (highlighted in white) indicates increased myelination upon Eotaxin-1 blockade. Scale bars: 100 μm (b), 50 μm (c,d), 500 μm (e). f, Schematic of behavioral test design: aged mice treated with Rat IgG or Eotaxin-1 antibody were tested for locomotor function and anxiety using the Open Field (OF) test, and their memory was assessed using the novel object recognition test (NOR) test. g, Time (sec) and distance spent (m) in the center or periphery of the arena during the OF test. h, Discrimination index, exploration time (sec) and total distance travelled (m) in the NOR test (n = 7/control; 10/Eotaxin-1 antibody). All data represented as mean ± s.e.m.; * P < 0.05, ** P < 0.01, Student’s t -test.

Article Snippet: For overexpression experiments, a separate cohort of 23-month-old C57BL/6J mice was treated with antibiotics for 30 days (as above) and concurrently injected intraperitoneally with recombinant mouse Eotaxin (10 ng/g body weight in PBS (2 μg/ml stock; R&D Systems, 420ME020) every 3 days.

Techniques: Control, Immunostaining, Labeling, Inhibition, Immunofluorescence, Activation Assay

Journal: bioRxiv

Article Title: Microbiome depletion rejuvenates the aging brain

doi: 10.64898/2026.02.13.705770

Figure Lengend Snippet: a, Flow cytometry analysis of peripheral blood from aged mice showing the frequency of major immune cell populations, including eosinophils, following Abx treatment. b, Representative images of CD170+ eosinophils (green) in the intestinal epithelium of young, aged control, and aged Abx-treated mice. c, Quantification of CD170+ eosinophils normalized to intestinal villus area in young, aged control, and aged Abx-treated mice (n = 4/group). d, Quantification of CD170+ eosinophils in the intestinal villi of aged mice treated with water + PBS or Abx + Eotaxin-1 (n = 3 control, n = 4 Abx + Eotaxin). White arrows indicate CD170 positive cells. Scale bar 100 µm. All data represented as mean ± s.e.m.; * P < 0.05, ** P < 0.01. ANOVA with Tukey’s multiple-comparisons post hoc test (c), Student’s t -test (d).

Article Snippet: For overexpression experiments, a separate cohort of 23-month-old C57BL/6J mice was treated with antibiotics for 30 days (as above) and concurrently injected intraperitoneally with recombinant mouse Eotaxin (10 ng/g body weight in PBS (2 μg/ml stock; R&D Systems, 420ME020) every 3 days.

Techniques: Flow Cytometry, Control

Journal: mBio

Article Title: Host cysteine proteases promote the severity of catheter-associated urinary tract infection and kidney fibrosis

doi: 10.1128/mbio.02161-25

Figure Lengend Snippet: Eosinophils play a protective role in CAUTI. At 2 dpi, single cells were isolated from the bladder mucosa of CAUTI mice. The percentages of eosinophils and neutrophils were measured by flow cytometry to assess the effects of E64 treatment ( A ) or eotaxin ( B ) as compared to mock. Total CFU recovered 2 dpi were quantified for catheter, bladder, and kidney for treatment by eotaxin ( C–E ), by a-IL-5 ( F–H ), or by CCR3 antagonist SB328437 ( I–K ). Each symbol represents an individual mouse, and symbols touching the dashed lines indicate values below the limit of detection (LOD, 40 CFU). Horizontal lines indicate mean values; error bars represent SEM. All data are from ≥3 biological replicates. Statistical comparisons were performed using a two-tailed unpaired Student’s t-test. * P < 0.05. Normality was assessed using the Shapiro-Wilk test.

Article Snippet: For drug therapy, E64 (Roche, 10874523001), anti-mouse IL-5 (BD Pharmingen, 554393), SB 328437 (CCR3 antagonist, Sigma, 247580-43-4), and mouse eotaxin (R&D systems, 420-ME) were dissolved in PBS according to the manufacturers’ recommendations to achieve the concentration stated in the text.

Techniques: Isolation, Flow Cytometry, Two Tailed Test