met ccl5  (R&D Systems)

Journal: Clinical Science (London, England : 1979)

Article Title: CCL5 deficiency aggravates acute DSS-induced colitis by restricting IL-33-induced formation of Tregs in intestinal tract

doi: 10.1042/CS20256734

Figure Lengend Snippet: Wildtype (WT) and CCL5 knockout ( Ccl5 -KO) mice of both C57BL/6 and BalB/C backgrounds were subjected to 7-day 2.5% DSS administration in the drinking water. ( A ) Schematic of construction of DSS-induced colitis model. ( B ) Disease activity index (DAI), n = 25 per group. ( C ) Survival rate of WT ( n = 25) and Ccl5 -KO ( n = 25) mice was observed for 2 weeks, and the corresponding survival curves were plotted. ( D and E ) Gross anatomy of colons ( D ) and colon length ( E ) were monitored. Colon length was measured in both the C57BL/6 and BalB/C mouse strains after 7 days of 2.5% DSS treatment. The results are shown as the mean ± SEM, with statistical significance assessed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. ( F and G ) Representative H&E staining ( F ) of colon sections from WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration (scale bars: 50 μm, 100 μm). Histological score ( G ) was quantified. Results shown are the mean ± SEM (ns, nonsignificant; ** P <0.01, *** P <0.001, **** P <0.0001) of triplicate determination from three independent experiments.

Article Snippet: To rescue the phenotypes of enteritis, exogenous recombinant murine CCL5 protein (HY- P71890 , MCE) [ ] was intraperitoneally injected at a concentration of 350 ng/kg from the onset of DSS treatment.

Techniques: Knock-Out, Activity Assay, Staining

Journal: Clinical Science (London, England : 1979)

Article Title: CCL5 deficiency aggravates acute DSS-induced colitis by restricting IL-33-induced formation of Tregs in intestinal tract

doi: 10.1042/CS20256734

Figure Lengend Snippet: ( A and B ) Flow cytometric analysis of number of CD4 + and CD8 + T cells in intestinal lymphoid tissues from mice with indicated genotypes 4 days after 7-day 2.5% dextran sulfate sodium salt (DSS) administration. Graphs ( B ) show the relative percentage of, respectively, CD4 + and CD8 + T cells ( n = 6 per group). ( C and D ) Representative CD4 staining of distal colon sections from wildtype (WT) and CCL5 knockout ( Ccl5 -KO) mice 4 days after 7-day 2.5% DSS administration ( C ; scale bars, 20 μm), with quantitative analysis ( D , n = 50); black arrows indicate CD4-positive cells. ( E and F ) Flow cytometric plots ( E ) of FOXP3 + CD4 + T cell population in intestines from control or DSS-treated mice with indicated genotypes ( n = 6 per group). Percentage of FOXP3 + population among CD4 + T cells is shown ( F , n = 6). ( G and H ) Representative FOXP3 staining of distal colon sections from WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration ( G ; scale bars: 10 μm, 50 μm), with quantitative analysis ( H , n = 50); red arrows indicate FOXP3-positive cells. ( I ) Immunoblotting of FOXP3 expression in colonic tissues from WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration. Results shown are the mean ± SEM (ns, nonsignificant; * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001) of triplicate determination from three independent experiments.

Article Snippet: To rescue the phenotypes of enteritis, exogenous recombinant murine CCL5 protein (HY- P71890 , MCE) [ ] was intraperitoneally injected at a concentration of 350 ng/kg from the onset of DSS treatment.

Techniques: Staining, Knock-Out, Control, Western Blot, Expressing

Journal: Clinical Science (London, England : 1979)

Article Title: CCL5 deficiency aggravates acute DSS-induced colitis by restricting IL-33-induced formation of Tregs in intestinal tract

doi: 10.1042/CS20256734

Figure Lengend Snippet: ( A ) Enzyme-linked immunosorbent assay (ELISA) analysis of the level of IL-33 in colon explant cultures at the indicated time points (days 0, 2, 4, 6, 8) during the 7-day dextran sulfate sodium salt (DSS) treatment in wildtype (WT) and CCL5 knockout ( Ccl5 -KO) mice ( n = 6 per group). ( B ) Immunoblotting of IL-33 and ST2 in intestinal epithelial cells of WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration. ( C ) Immunoblotting of NF-κB ( P65 ) related pathway in intestinal epithelium of WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration. ( D ) Immunofluorescence staining for P65 (red) in colonic sections of WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration; DNA (DAPI, blue); scale bars, 50 μm. White arrows indicate P65-positive cells with nuclear translocation. ( E ) Immunoblotting of PI3K/Akt pathway in intestinal epithelium of WT and Ccl5 -KO mice 4 days after 7-day 2.5% DSS administration. ( F ) ELISA analysis of IL-33 levels in colon explant cultures of WT and Ccl5 -KO mice at the indicated time points (days 0, 2, 4, 6, 8) during 7-day DSS treatment with CCL5 small protein interventions ( n = 6 per group). ( G ) Colon length measurements on day 12 in Ccl5 -KO mice treated with different drug groups (control, CCL5 small protein, CCL5 small protein + BAY 11-7082, CCL5 small protein + Capivasertib); n = 6 per group. ( H ) Recording of DAI different time points ( D0, D3, D6, D9, D12 ) in different drug treatment groups during the treatment period. ( I ) Immunoblot analysis of corresponding protein levels in the intestinal epithelial tissues of Ccl5 -KO mice after treatment with different drug groups. ( J ) Immunofluorescence staining analysis of P65-positive (red) cells in the intestines of Ccl5 -KO mice treated with different drug groups (control, CCL5 small protein, CCL5 small protein + Capivasertib); DNA (DAPI, blue), scale bars, 50 μm. White arrows indicate P65-positive cells with nuclear translocation. Results shown are the mean ± SEM (ns, nonsignificant; * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001) of triplicate determination from three independent experiments.

Article Snippet: To rescue the phenotypes of enteritis, exogenous recombinant murine CCL5 protein (HY- P71890 , MCE) [ ] was intraperitoneally injected at a concentration of 350 ng/kg from the onset of DSS treatment.

Techniques: Enzyme-linked Immunosorbent Assay, Knock-Out, Western Blot, Immunofluorescence, Staining, Translocation Assay, Control

Journal: Clinical Science (London, England : 1979)

Article Title: CCL5 deficiency aggravates acute DSS-induced colitis by restricting IL-33-induced formation of Tregs in intestinal tract

doi: 10.1042/CS20256734

Figure Lengend Snippet: ( A ) Immunoblotting of IL-33, ST2, forkhead box protein 3 (FOXP3), and JAK1/STAT5 signaling in intestinal CD4 + T cells of wildtype (WT) and CCL5 knockout ( Ccl5 -KO) mice 4 days after 7-day 2.5% dextran sulfate sodium salt (DSS) administration. ( B and C ) Mice received daily intraperitoneal injections of rIL-33 protein (10 ng/µL) at the onset of the DSS regimen. The intraperitoneal administration of different therapeutic agents continued for an additional 4 days after 7-day DSS treatment. Subsequently, the mice were killed, and the affected intestinal tissues were analyzed. Disease activity index (DAI) ( B ) and colon length ( C ) were monitored, n = 6 per group. ( D and E ) Representative H&E staining ( D ) of colon sections in mice from different treated groups (scale bars, 50 μm). Histological score ( E ) was quantified. ( F and G ) Representative FOXP3 staining of distal colon sections from rescued DSS-treated mice with indicated genotypes after 8-day rIL-33/PBS treatment ( F ; scale bars, 20 μm) and quantitative analysis ( G , n = 50). ( H and I ) Flow cytometric plots ( H ) of FOXP3 + CD4 + T cell population in intestines from rescued DSS-treated mice with indicated genotypes after 8-day rIL-33/PBS treatment and quantitative analysis ( I , n = 6). ( J ) Immunoblotting of FOXP3 and JAK1/STAT5 signaling in intestinal CD4 + T cells, respectively, from rescued DSS-treated mice with indicated genotypes after 8-day rIL-33/PBS treatment. The results are shown as the mean ± SEM (ns, nonsignificant; * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001) of triplicate determination from three independent experiments, with statistical significance assessed using one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test.

Article Snippet: To rescue the phenotypes of enteritis, exogenous recombinant murine CCL5 protein (HY- P71890 , MCE) [ ] was intraperitoneally injected at a concentration of 350 ng/kg from the onset of DSS treatment.

Techniques: Western Blot, Knock-Out, Activity Assay, Staining

Journal: Clinical Science (London, England : 1979)

Article Title: CCL5 deficiency aggravates acute DSS-induced colitis by restricting IL-33-induced formation of Tregs in intestinal tract

doi: 10.1042/CS20256734

Figure Lengend Snippet: ( A ) Immunoblotting ( n = 12) of CCL5 and FOXP3 expression in inflamed tissue in ulcerative colitis (UC) patients. ( B ) Quantitative polymerase chain reaction (qPCR) analysis ( n = 32) of CCL5 expression, respectively, in intestinal epithelial (IECs) and stromal cells (SCs) from inflamed and adjacent normal tissue in UC patients. ( C and D ) Representative H&E staining (right), CCL5 (middle), and FOXP3 (left) staining of inflamed intestinal tissue from UC patients with different levels of CCL5 expression (CCL5 [high] vs. CCL5 [low]), ( C ; scale bars, 20 μm, 100 μm) and quantitative analysis of CCL5 and FOXP3 expression ( D , n = 32 per group) in inflamed (UC) and adjacent normal tissue (NC). ( E ) Correlation analysis between the proportion of CCL5-positive cells and FOXP3-positive cells in inflamed (UC) tissue from UC patients ( n = 32). ( F ) Quantitative real-time polymerase chain reaction (RT-qPCR) analysis of the correlation between CCL5 and FOXP3 expression levels in inflamed intestinal tissue ( n = 32) from UC patients with different levels of CCL5 expression. ( G ) Flow cytometric plots (right) of FOXP3 + CD4 + T cell population in intestines from CCL5-low and CCL5-high UC patient tissues with quantitative analysis (left, n = 6). Results shown are the mean ± SEM (ns, nonsignificant; *** P <0.001, **** P <0.0001) of triplicate determination from three independent experiments.

Article Snippet: To rescue the phenotypes of enteritis, exogenous recombinant murine CCL5 protein (HY- P71890 , MCE) [ ] was intraperitoneally injected at a concentration of 350 ng/kg from the onset of DSS treatment.

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Staining, Quantitative RT-PCR

Journal: Biomedicines

Article Title: The Chemokine (C-C Motif) Receptor 1 Antagonist BX471 Improves Fluid Resuscitation in Rat Models of Hemorrhagic Shock

doi: 10.3390/biomedicines13051241

Figure Lengend Snippet: Serum CCR1 ligand concentrations (pg/mg) at baseline (t = 0 min) and at the end of the experiment (t = 210 min). Light grey bars: vehicle treatment, n = 7. Dark grey bars: 0.5 μmol/kg BX471 treatment, n = 7. ( A ) CCL3, ( B ) CCL4, ( C ) CCL5, and ( D ) CCL7. *: p < 0.05 vs. baseline (t = 0 min).

Article Snippet: Measurements of Chemokine Concentrations: Serum levels of CCL3, CCL4, CCL5, and CCL7 were measured with commercially available rat enzyme-linked immunosorbent assays (ELISA, Boster Bio, Pleasanton, CA, USA) according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Immunology

Article Title: NOX4 modulates breast cancer progression through cancer cell metabolic reprogramming and CD8 + T cell antitumor activity

doi: 10.3389/fimmu.2025.1534936

Figure Lengend Snippet: NOX4 enhanced the CD8 + T cells mediate antitumor effect. (A) TIMER2.0 was used to analysis the correlation between NOX4 expression levels and CD8 + T cell infiltration, Scatter plot showing the correlation between NOX4 expression levels and CD8 + T cell infiltration in breast cancer (BRCA) samples (n=1100). Rho=0.346, p=2.57e-26. (B) TISIDB data: Scatter plot showing the correlation between NOX4 expression and central memory CD8 + T cell (Tcm-CD8) infiltration, effector memory CD8+ T cell (Tem-CD8) in BRCA samples (n=1100). Rho=0.44, p<2.2e-16; Rho=0.171, p<1.24e-6. (C) Representative flow cytometry plots showing the percentage of CD8 + T cells within the CD45 + population in EO771 tumors from NOX4 wild-type (WT) and NOX4 knockout (KO) mice. Bar graph quantifying the percentage of CD8 + cells, showing a significant reduction in CD8 + T cell infiltration in NOX4 KO tumors ***p<0.001. n=6 per group. (D) Representative flow cytometry plots showing IFN-γ and Granzyme B expression in CD8 + T cells from EO771 tumors of NOX4 WT and NOX4 KO mice. Bar graphs quantifying the percentage of CD8 + T cells expressing IFN-γ and Granzyme B, ***p<0.001. n=7 per group. (E) Representative flow cytometry plots showing the percentage of DC cells within the EO771 tumors from NOX4 wild-type (WT) and NOX4 knockout (KO) mice. Bar graph quantifying the percentage of DC cells, **p<0.01. n=7 per group. (F) TISIDB data: Scatter plot showing the correlation between NOX4 expression and CCL11 expression in BRCA samples (n=1100). A significant positive correlation is observed. Rho=0.359, p<2.2e-16. Bar graph showing the concentration of CCL11, CCL5 in the tumor microenvironment, *p<0.05. Data are presented as mean ± SEM from 3 independent experiments. Statistical significance was determined using two-tailed Student’s t-test.

Article Snippet: The quantity of CCL11, CCL5 (Bosterbio) were determined in tumor tissue using ELISA kits according to the manufacturer’s instructions.

Techniques: Expressing, Flow Cytometry, Knock-Out, Concentration Assay, Two Tailed Test