recombinant 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: Journal of Neuroinflammation

Article Title: Astrocytic CCL5 orchestrates CCR5-positive neuronal necroptosis in subarachnoid hemorrhage

doi: 10.1186/s12974-026-03714-5

Figure Lengend Snippet: Astrocytic CCL5 is the primary ligand upregulated after SAH. A . Heatmap of chemokine mRNA expression 3 days post-SAH by qRT-PCR ( n = 3). B . ELISA of CCL5 protein levels in brain tissue at each indicated time ( n = 6). C . Regional CCL5 mRNA expression by qRT-PCR ( n = 5). D - E . Immunofluorescent staining showed that CCL5 was mainly colocalized with GFAP⁺ astrocytes, but not Iba1⁺ microglia, in the hippocampus. Scale bars, 25 μm. Data are mean ± SD; * P < 0.05, ** P < 0.01 vs. sham; one-way ANOVA with Tukey’s test

Article Snippet: Additionally, recombinant CCL5 (rCCL5, 0.5 μg/ mouse; R&D Systems, Minneapolis) was dissolved in PBS and administered to SAH mice via intracerebroventricular injection as previously reported [ ].

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Neuroinflammation

Article Title: Astrocytic CCL5 orchestrates CCR5-positive neuronal necroptosis in subarachnoid hemorrhage

doi: 10.1186/s12974-026-03714-5

Figure Lengend Snippet: Knockdown of astrocytic CCL5 reduces neuronal death and improves neurobehavioral function. A . Schematic timeline of the experimental protocol. B . Immunofluorescence images showed the expression of CCL5 and GFAP with shCCL5 or vector intervention. Scale bar: 50 μm. C . ELISA analysis of CCL5 levels in the hippocampus with shCCL5 or vector intervention ( n = 6), * P < 0.05 indicated comparison with vector group. D - E . TUNEL staining and quantification of neuronal death in the hippocampus at 3d after SAH ( n = 6), Scale bars, 25 μm. F – H . Western blot and quantification of PSD95 and Syn in the hippocampus at 3d after SAH ( n = 6). I–M. Neurobehavioral tests: Garcia score ( I ), Rotarod test ( J ), Morris water maze escape latency ( K ), distance to platform ( L ), and platform crossings ( M ) ( n = 6). Data are mean ± SD; * P < 0.05 and ** P < 0.01 indicated comparison with sham group. # P < 0.05 indicated comparison with SAH + vector group; one-way ANOVA with Tukey’s test

Article Snippet: Additionally, recombinant CCL5 (rCCL5, 0.5 μg/ mouse; R&D Systems, Minneapolis) was dissolved in PBS and administered to SAH mice via intracerebroventricular injection as previously reported [ ].

Techniques: Knockdown, Immunofluorescence, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Comparison, TUNEL Assay, Staining, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Astrocytic CCL5 orchestrates CCR5-positive neuronal necroptosis in subarachnoid hemorrhage

doi: 10.1186/s12974-026-03714-5

Figure Lengend Snippet: CCL5 and CCR5 levels in human CSF correlate with SAH severity and outcome. A . ELISA of CCR5 in CSF from control and SAH patients ( n = 22 control, n = 41 SAH). B . Correlation of CSF level of CCR5 with Hunt-Hess grade of SAH patients. C . CSF levels of CCR5 in patients with good (mRS 0–2) vs. poor (mRS 3–6) outcomes. D . ELISA of CCR5 in CSF from control and SAH patients. E . Correlation of CSF level of CCL5 with Hunt-Hess grade of SAH patients. F . CSF levels of CCL5 in patients with good vs. poor outcomes. Data are mean ± SD; P < 0.05; Student’s t-test or Pearson correlation ( B , E )

Article Snippet: Additionally, recombinant CCL5 (rCCL5, 0.5 μg/ mouse; R&D Systems, Minneapolis) was dissolved in PBS and administered to SAH mice via intracerebroventricular injection as previously reported [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cancer Research

Article Title: FADD Activation in Hepatocellular Carcinoma Potentiates CD8 + T-cell Responses and Sensitizes to Immune Checkpoint Inhibitors

doi: 10.1158/0008-5472.CAN-24-3854

Figure Lengend Snippet: FADD in hepatoma cells directs intratumoral CD8 + T-cell infiltration via the CCL5–CCR5 chemotaxis pathway. A, Schematic of the in vivo T-cell migration assay. B, Tumor weights and relative FADD mRNA levels in HepG2-vector and HepG2- FADD tumors 14 days after inoculation ( n = 8). C, Representative IHC images of CD8 in HepG2-vector and HepG2- FADD tumors and quantification thereof ( n = 10). Scale bar, 100 μm. D, Tumor weights and relative FADD mRNA levels in Huh7-sh Ctrl and Huh7-sh FADD tumors 14 days after inoculation ( n = 8). E, Representative IHC images of CD8 in Huh7-sh Ctrl and Huh7-sh FADD tumors and quantification thereof ( n = 10). Scale bar, 100 μm. F, Venn diagram of 521 immune-related genes from and cytokines/chemokines from ImmPort identified 36 overlapping cytokines and chemokines. G, mRNA levels of the 36 cytokines and chemokines in HepG2-vector, HepG2-FADD, Huh7-sh Ctrl , and Huh7-sh FADD cells, as well as tumors generated in A . H, Schematic of NSG-HuPBL humanized mouse model ( n = 8–10). I and J, Representative coimmunofluorescence images of CD8 and CCR5 in HepG2-vector and HepG2- FADD ( I ) and Huh7-sh Ctrl and Huh7-sh FADD tumors ( J ). Hoechst marks cell nuclei. Quantification of CCR5 + CD8 + cell proportions in the indicated groups is also provided. Scale bar, 100 μm. Data are presented as the mean ± SD for at least two (mouse models) or three (Western blotting) independent experiments and analyzed with the paired ( B–E , I and J ) or unpaired ( G ) two-tailed Student t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: HepG2- FADD or Huh7-sh FADD cells were then treated with CCL5-neutralizing antibodies (nAb; 100 pg/mL, R&D Systems, #MAB678-SP) or recombinant human CCL5 (rCCL5; 100 pg/mL, R&D Systems, #278-RN-050/CF), respectively, before T cells were added to the top chamber.

Techniques: Chemotaxis Assay, In Vivo, Cell Migration Assay, Plasmid Preparation, Generated, Western Blot, Two Tailed Test

Journal: Cancer Research

Article Title: FADD Activation in Hepatocellular Carcinoma Potentiates CD8 + T-cell Responses and Sensitizes to Immune Checkpoint Inhibitors

doi: 10.1158/0008-5472.CAN-24-3854

Figure Lengend Snippet: FADD upregulates CCL5 by activating NF-κB transcription in HCC cells. A, Western blotting analysis of p-FADD Ser194 , FADD, p-IκBα Ser32 , IκBα, NF-κB-p50, p-NF-κB-p65 Ser536 , and NF-κB-p65. B, Relative CCL5 mRNA levels in the indicated groups. C, Relative mRNA levels of FADD and CCL5 in HepG2-vector and HepG2-FADD cells treated with siNC, siNF-κB-p50, or siNF-κB-p65. D, ChIP-qPCR analysis of NF-κB-p65 in the CCL5 promoter of HepG2-vector, HepG2- FADD , and HepG2- FADD -S194A stable cells. The CCL5 promoter region for TF binding is also shown. Data are presented as the mean ± SD for at least three (Western blot and qPCR) or two (ChIP-qPCR) independent experiments and analyzed with the unpaired, two-tailed Student t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: HepG2- FADD or Huh7-sh FADD cells were then treated with CCL5-neutralizing antibodies (nAb; 100 pg/mL, R&D Systems, #MAB678-SP) or recombinant human CCL5 (rCCL5; 100 pg/mL, R&D Systems, #278-RN-050/CF), respectively, before T cells were added to the top chamber.

Techniques: Western Blot, Plasmid Preparation, ChIP-qPCR, Binding Assay, Two Tailed Test

Journal: Cancer Research

Article Title: FADD Activation in Hepatocellular Carcinoma Potentiates CD8 + T-cell Responses and Sensitizes to Immune Checkpoint Inhibitors

doi: 10.1158/0008-5472.CAN-24-3854

Figure Lengend Snippet: Phosphorylated FADD promotes NF-κB transcriptional activity via formation of a nuclear p-FADD/SAM68/NF-κB complex in HCC cells. A, Representative co-immunofluorescence images of p-FADD Ser194 (red), FADD (green), and nuclei (DAPI, blue) in HepG2- FADD and HepG2-FADD-S194 stable cell lines. Scale bar, 100 μm. B, Western blotting analysis of p-FADD Ser194 and FADD in the cytoplasmic (Cyto) and nuclear (Nuc) fractions. C, Coomassie brilliant blue staining for co-IP of nuclear proteins pulled down with anti-FADD antibody in HepG2-vector, HepG2-FADD, and HepG2-FADD-S194A stable cell lines. D, Venn diagram of 547 proteins identified through MS analysis in samples from C . E, Correlations of the 110 unique HepG2- FADD nuclear proteins with FADD expression, positive regulation of IκB kinase/NF-κB signaling pathway signatures, and positive regulation of NF-κB TF activity in samples from the TCGA-LIHC dataset. F, Protein–protein interaction network analysis of SAM68 with eight nodes. An average node degree of 4.25 with a median (0.4) level of confidence is required as the minimum required interaction score. G, Co-IP of nuclear FADD and SAM68 in HepG2-vector, HepG2-FADD, and HepG2-FADD-S194A stable cell lines, followed by Western blotting analysis of p-FADD Ser194 , FADD, SAM68, NF-κB-p50, p-NF-κB-p65 Ser536 , and NF-κB-p65. Lamin B serves as the loading control for nuclear lysate input, and IgG is the control for the IP assay. L, ladder. H, Western blotting analysis of SAM68, p-FADD Ser194 , FADD, p-IκBα Ser32 , IκBα, NF-κB-p50, p-NF-κB-p65 Ser536 , and NF-κB-p65. I, Relative mRNA levels of FADD and CCL5 in HepG2-vector and HepG2- FADD cells treated with siNC or siSAM68 (#1 and #2). J, Co-IP of nuclear FADD and SAM68 from HepG2-vector, HepG2- FADD , and HepG2-FADD treated with siSAM68 (#2), followed by Western blotting analysis of p-FADD Ser194 , FADD, SAM68, NF-κB-p50, p-NF-κB-p65 Ser536 , and NF-κB-p65. Histone H3 served as the nuclear lysate input loading control, and IgG is the IP assay control. K, Relative induction of NF-κB luciferase in HepG2-vector and HepG2- FADD cells treated with siNC or siSAM68 (#2). Data are presented as the mean ± SD for at least three independent experiments (coimmunofluorescence, Western blotting, and co-IP) and analyzed with the unpaired, two-tailed Student t test. Two-tailed Pearson correlation was used to compute the correlation between variables. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: HepG2- FADD or Huh7-sh FADD cells were then treated with CCL5-neutralizing antibodies (nAb; 100 pg/mL, R&D Systems, #MAB678-SP) or recombinant human CCL5 (rCCL5; 100 pg/mL, R&D Systems, #278-RN-050/CF), respectively, before T cells were added to the top chamber.

Techniques: Activity Assay, Immunofluorescence, Stable Transfection, Western Blot, Staining, Co-Immunoprecipitation Assay, Plasmid Preparation, Expressing, Control, Luciferase, Two Tailed Test

Journal: Cancer Research

Article Title: FADD Activation in Hepatocellular Carcinoma Potentiates CD8 + T-cell Responses and Sensitizes to Immune Checkpoint Inhibitors

doi: 10.1158/0008-5472.CAN-24-3854

Figure Lengend Snippet: Sequential activation of FADD converts anti–PD-1 responsiveness in ICI-resistant HCC mouse models. A, Schematic of sequential anti–PD-1 treatment and Fadd overexpression in orthotopic PD-1R tumor–bearing C57BL/6 mice. B, Kaplan–Meier survival analysis of mice from the indicated groups ( n = 8–10). C and D, Western blotting analysis of p-Fadd Ser191 and Fadd in tumors from control or AAV- Fadd –treated tumor-bearing mice ( C ) and CTNNB1 OE /MYC OE -induced spontaneous tumors ( D ). E, Schematic of sequential anti–PD-1 and ADT-OH treatment schedule in the CTNNB1 OE /MYC OE -induced spontaneous HCC model. F, Western blotting analysis of p-Fadd Ser191 and Fadd in tumors from control and ADT-OH–treated mice. G and H, Representative photos, hematoxylin and eosin staining ( G ), and enumeration of tumor nodules in the indicated groups on day 80 ( H ). Scale bar, 1,000 μm. I, Uniform Manifold Approximation and Projection (UMAP) plots showing high-dimensional flow cytometry analysis of tumor-infiltrating CD45 + leukocytes from the indicated groups of mice ( n = 6). UMAP plots consist of 12,000 cells each and are representative of concatenated samples within each group. Indicated immune cell clusters are highlighted in the indicated colors, and their proportions as percentages of CD45 + leukocytes are quantified ( J ). DC, dendritic cell; NA, no annotation. K, Heat map showing Pearson correlations among the number of tumor nodules; p-Fadd Ser191 ; Fadd; CCL5; the proportions of total, CCR5 + , IFNγ + , and TNFα + CD8 + T cells; TUNEL scores; and the concentrations of IFNγ and TNFα. L, Kaplan–Meier survival analysis of mice from the indicated groups ( n = 10–14); two-sided log-rank (Mantel–Cox) test. M, The FADD/CCL5/CD8a signature in patients with HCC treated with atezolizumab alone or in combination with bevacizumab (anti-VEGF). Data are presented as the mean ± SD for at least two (mouse models) or three (Western blotting) independent experiments and analyzed with the unpaired, two-tailed Student t test. Correlation analyses were performed using single-tailed Pearson correlations. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: HepG2- FADD or Huh7-sh FADD cells were then treated with CCL5-neutralizing antibodies (nAb; 100 pg/mL, R&D Systems, #MAB678-SP) or recombinant human CCL5 (rCCL5; 100 pg/mL, R&D Systems, #278-RN-050/CF), respectively, before T cells were added to the top chamber.

Techniques: Activation Assay, Over Expression, Western Blot, Control, Staining, Flow Cytometry, TUNEL Assay, Two Tailed Test