elisa kit  (R&D Systems)

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A Multiplexing analysis of 10 post-operative circulating cytokines in HCC patients who received liver resection. Non-Recur, non-recurrence ( N = 22); Recur, Recurrence ( N = 26). B Comparison of post-resection serum CCL11 protein between patients with and without HCC recurrence after hepatectomy. C ROC analysis of post-resection serum CCL11 protein in distinguishing patients with HCC recurrence after hepatectomy. D Kaplan-Meier analysis of post-resection serum CCL11 protein (High group vs Low group) in association with patients’ disease-free survival. E Nomogram of CCL11-integrated prediction model in predicting 5-year HCC recurrence after hepatectomy. F Comparison of ROC curves of CCL11-integrated predictive model with reference model (without CCL11) in predicting 5-year HCC recurrence after hepatectomy. G The expression level of CCL11mRNA in tumor and non-tumor liver tissues of HCC patients as well as in healthy liver tissues. H Comparison of the expression level of non-tumoral CCL11 (ntCCL11) mRNA between patients with and without HCC recurrence. I ROC curves of ntCCL11 mRNA levels in predicting 5-Year HCC recurrence after hepatectomy. J Kaplan-Meier analysis of ntCCL11 mRNA in predicting disease-free survival. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns not significant.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques: Multiplexing, Comparison, Expressing

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A Correlation analyses of CCL11 levels with its receptors CCR2, CCR3 and CCR5 in non-tumoral liver tissue. B UMAP plot of CCR5+immune cells in CCL11-low group and CCL11-high group. C UMAP plot of all immune cell types. D Correlations of CCL11 levels with iNOS and CD206 in non-tumor liver tissues. E Correlation of CCL11 with CCR5 + M2-like macrophages. Kaplan-Meier survival curve of ( F ) overall survival and G disease-free survival based on CCR5 + M2-like macrophages. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques:

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A Flow cytometry quantification of the number of monocyte-differentiated CCR5 + CD206 + M2-like macrophages after administration of recombinant human CCL11 (rhCCL11) protein. Control group: PBS treatment. B The expression Levels of M2-macrophage-related immunosuppressive markers and cytokines in CCL11-treated and PBS-treated CCR5 + M2-like macrophages. C Cytokine array analysis of secreted cytokines in CCL11-treated and PBS-treated CCR5 + M2-like macrophages. Red dot boxes indicate two highly upregulated cytokines: 1, MCP1; 2, IL-8. D mRNA Levels of IL-8 and MCP-1 in CCL11-treated and PBS-treated CCR5 + M2-like macrophages. E CCL11-treated CCR5 + M2-like macrophages enhanced the induction of CD4+ Foxp3+Tregs from naive CD4 + T compared to the PBS-treated control group. F Top 10 enriched KEGG pathways from differentially expressed genes in the CCL11-treated CCR5 + M2-like macrophages. G Gene set enrichment analysis of Toll-like receptor signaling pathway. H Levels of p-IKKα/β, p-IkBα, NFkB1-P50, and PD-L1 expression via western blot analysis. I Schematic diagram of the molecular mechanism of CCL11 in regulating CCR5 + M2-like macrophages. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns not significant.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques: Flow Cytometry, Recombinant, Control, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A Wound-healing assay to evaluate the migration of Hep3B with different concentrations of recombinant CCL11 protein (0, 12.5, 25, 50 ng/ml). B Wound-healing assay of Hep3B when blockage of CCL11-receptor CCR2, CCR3 and CCR5 in CCL11-enriched condition; C Transwell migration assay of Hep3B when blockage of CCL11-receptor CCR2, CCR3 and CCR5 in CCL11-enriched condition. D Transwell invasion assay of Hep3B when blockage of CCL11-receptor CCR2, CCR3 and CCR5 in CCL11-enriched condition. Kaplan-Meier survival curve of ( E ) Overall survival and F disease-free survival based on the expression level of CCR3 mRNA in HCC patients. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns not significant.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques: Wound Healing Assay, Migration, Recombinant, Transwell Migration Assay, Transwell Invasion Assay, Expressing

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A RT2 PCR array analysis of 84 metastasis-associated genes in CCL11-treated and PBS-treated (Ctrl) HCC cells. B RT2 PCR array analysis of 84 metastasis-associated genes in CCL11 treatment (50 ng/ml) with anti-CCR3, compared to CCL11 single treatment. C Identification of upregulated genes in CCL11-high and CCR3-high in HCC from TCGA database. D Top 20 KEGG pathways associated with tumor metastasis among the 92 upregulated genes in CCL11-high & CCR3-high group. E The expression level of MMP13 mRNA CCL11-treated HCC cells followed by administration of LY294002 (PI3K inhibitor), compared with DMSO-treated control cells. F Migration assay and G Invasion assay of CCL11-treated HCC cells with or without adding LY294002. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques: Expressing, Control, Migration, Invasion Assay

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A The bioinformatic analysis of MMP13-promoter binding transcript factor and AKT-activating transcript factor. B The promoter region of MMP13 gene contains 2 binding motifs for the AKT downstream transcription factor Mafk, located at nucleotide sequences -232 to -218 and -57 to -43. C Inhibition of PI3K significantly reduced the CCL11-induced Mafk transloation from cytoplasm to nuclear in HCC cells. D Western blot analysis of p-PI3K, PI3K, p-Akt, Akt, Mafk, and MMP13 protein in CCL11-treated HCC cells with and without PI3K inhibition. E Schematic diagram of mouse orthotopic HCC tumor model, followed by injection of rCCL11 Protein or PBS via portal vein. F IVIS evaluations of orthotopic HCC tumor in rCCL11 Protein group or PBS group. G Liver organs dissected from rCCL11 group or PBS group. H Western blot analysis of PI3K/Akt/MafK/MMP13 axis in rCCL11 protein group and PBS group. I Schematic diagram of the molecular mechanism of CCL11 in promoting HCC progression. *, P < 0.05; **, P < 0.01.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques: Binding Assay, Inhibition, Western Blot, Injection

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A Schematic diagram of orthotopic HCC resection model, followed by anti-CCL11 therapy. B IVIS evaluations of intrahepatic and extrahepatic tumors in the control group and anti-CCL11 treatment group. C Recurred rates of recurred tumors in the control group and anti-CCL11treatment group. D Kaplan-Meier analysis for overall survival of mice after treatment. E Summary of molecular mechanisms of CCL11 in promoting HCC recurrence after liver resection. *, P < 0.05.

Article Snippet: Similarly, mouse serum CCL11 levels were measured with the Mouse CCL11/Eotaxin ELISA Kit (Proteintech).

Techniques: Control

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