Journal: Aging

Article Title: Targeting PRKDC activates the efficacy of antitumor immunity while sensitizing to chemotherapy and targeted therapy in liver hepatocellular carcinoma.

doi: 10.18632/aging.205855

Figure Lengend Snippet: Figure 9. PRKDC knockdown sensitizes Huh-7 and Hep-3B to chemotherapeutic and immunotherapeutic agents. (A) Dose response curves of Gemcitabine. (B) Dose response curves of Sorafenib. (C, D) Chemokines including CCL2, CCL4, CCL5, CLCX1, CLCX8 and CLCX10 are detected using ELISA and qPCR. (E, F) Schematic representation of the CD8+ T cell migration experiment, along with the comparative migration of CD8+ T cells between the PRKDC knockdown and the control groups. (G) Dose response curves of IFN-γ in Huh-7 and Hep-3B co-cultured with CD8+ T cells. (H, I) Comparison of T cell-induced tumor cell death between PRKDC-silenced and control groups in Huh-7 or Hep-3B cells. (*P < 0.05, **P < 0.01, and ***P < 0.001 determined by two-way ANOVA test).

Article Snippet: Supernatants from cell cultures in 24-well plates were harvested, and the chemokines including CCL2, CCL4, CCL5, CXCL1, CXCL8, and CXCL10 were identified with ELISA kit (Proteintech, USA).

Techniques: Knockdown, Enzyme-linked Immunosorbent Assay, Migration, Control, Cell Culture, Comparison

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Identification of Heparin-binding Sites in Proteins by Selective Labeling *

doi: 10.1074/mcp.M900031-MCP200

Figure Lengend Snippet: HBS of protect and label strategy test proteins The HBSs and the methods used to identify these are given. SDM, site-directed mutagenesis; x-ray, x-ray crystallography; SP, synthetic peptide; MM, molecular modeling.

Article Snippet: Platelet factor-4 (PF-4) (Swiss-Prot accession number P02776 ) (residues 32–101) was purchased from Athens Research and Technology Inc. (Athens, GA).

Techniques: Mutagenesis, Structural Proteomics

Journal: PLoS ONE

Article Title: SNPs in inflammatory genes CCL11 , CCL4 and MEFV in a fibromyalgia family study

doi: 10.1371/journal.pone.0198625

Figure Lengend Snippet: SNPs of immune related genes in chromosome 17 from FM cohorts and transmission analysis from FM trios ¹ , ² , ³ .

Article Snippet: Recombinant human CCL3 (MIP-1 α ), CCL4 (MIP-1 β ), CCL11 (Eotaxin), CCL1 (MCP-1), IL-13, GM-CSF, IL-4, and IFN γ were obtained from ProSpec Protein-Specialists (East Brunswick, NJ, USA); LPS from E. coli (055:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA); recombinant human CCL4 variant (S80T) was from Kingfisher Biotech, Inc (St Paul, MN, USA); anti-human CCR5-PE (CD195, Clone# J418F1), anti-human CCR1-APC (CD191, Clone# 5F10B29), anti-human CD4-PerCP (Clone# OKT4) and anti-human CD3-FITC (clone# SK7) were from Biolegend (San Diego, CA, USA); Indo-1-AM was from Molecular Probes (Eugene, OR).

Techniques: Transmission Assay

Journal: PLoS ONE

Article Title: SNPs in inflammatory genes CCL11 , CCL4 and MEFV in a fibromyalgia family study

doi: 10.1371/journal.pone.0198625

Figure Lengend Snippet: A-C , CCL11, CCL4, and CCL3 levels in female healthy controls (Ctrl, n = 48) vs female FM patients with wild type CCL11 (n = 19) or var CCL11 (n = 14). D-F , CCL4, CCL3 and CCL11 levels in female healthy controls (Ctrl, n = 48) vs wild type CCL4 (n = 49) or varCCL4 (n = 24) in female FM patients. (* P<0.05, ** P<0.01, *** P<0.001 in comparison with female healthy controls; # P<0.05 in comparison with female FM patients with wild type CCL4.

Article Snippet: Recombinant human CCL3 (MIP-1 α ), CCL4 (MIP-1 β ), CCL11 (Eotaxin), CCL1 (MCP-1), IL-13, GM-CSF, IL-4, and IFN γ were obtained from ProSpec Protein-Specialists (East Brunswick, NJ, USA); LPS from E. coli (055:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA); recombinant human CCL4 variant (S80T) was from Kingfisher Biotech, Inc (St Paul, MN, USA); anti-human CCR5-PE (CD195, Clone# J418F1), anti-human CCR1-APC (CD191, Clone# 5F10B29), anti-human CD4-PerCP (Clone# OKT4) and anti-human CD3-FITC (clone# SK7) were from Biolegend (San Diego, CA, USA); Indo-1-AM was from Molecular Probes (Eugene, OR).

Techniques: Comparison

Journal: PLoS ONE

Article Title: SNPs in inflammatory genes CCL11 , CCL4 and MEFV in a fibromyalgia family study

doi: 10.1371/journal.pone.0198625

Figure Lengend Snippet: A-B . WT CCL4 and varCCL4 both stimulate and cross-block calcium mobilization in CCR5 positive immature dendritic cell ( A ) or CEM.NK R CCR5 T-cells ( B ). Ionomycin and CCL3 were used as positive controls. C-D . varCCL4 induces greater down-regulation of cell surface CCR5 than WT CCL4 on immature dendritic cells ( C ) and CEM.NK R CCR5 T-cell line ( D ). Cells were treated with 80 ng/mL of each CCL4 species for 3 hours, surface CCR5 was stained with anti-CCR5 antibody and analyzed by flow cytometry (** P<0.01, *** P<0.001 in comparison with untreated control; # P<0.05 in comparison with WT CCL4).

Article Snippet: Recombinant human CCL3 (MIP-1 α ), CCL4 (MIP-1 β ), CCL11 (Eotaxin), CCL1 (MCP-1), IL-13, GM-CSF, IL-4, and IFN γ were obtained from ProSpec Protein-Specialists (East Brunswick, NJ, USA); LPS from E. coli (055:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA); recombinant human CCL4 variant (S80T) was from Kingfisher Biotech, Inc (St Paul, MN, USA); anti-human CCR5-PE (CD195, Clone# J418F1), anti-human CCR1-APC (CD191, Clone# 5F10B29), anti-human CD4-PerCP (Clone# OKT4) and anti-human CD3-FITC (clone# SK7) were from Biolegend (San Diego, CA, USA); Indo-1-AM was from Molecular Probes (Eugene, OR).

Techniques: Blocking Assay, Staining, Flow Cytometry, Comparison, Control

Journal: PLoS ONE

Article Title: SNPs in inflammatory genes CCL11 , CCL4 and MEFV in a fibromyalgia family study

doi: 10.1371/journal.pone.0198625

Figure Lengend Snippet: Purified CD4 T-cells from mismatched donors were co-incubated for nine days and treated with WT or varCCL4 (50ng/mL) for 3 hours, surface stained with anti-CCR5 antibody and analyzed by flow cytometry (** P<0.01, *** P<0.001 in comparison with untreated control; # P<0.05 in comparison with WT CCL4).

Article Snippet: Recombinant human CCL3 (MIP-1 α ), CCL4 (MIP-1 β ), CCL11 (Eotaxin), CCL1 (MCP-1), IL-13, GM-CSF, IL-4, and IFN γ were obtained from ProSpec Protein-Specialists (East Brunswick, NJ, USA); LPS from E. coli (055:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA); recombinant human CCL4 variant (S80T) was from Kingfisher Biotech, Inc (St Paul, MN, USA); anti-human CCR5-PE (CD195, Clone# J418F1), anti-human CCR1-APC (CD191, Clone# 5F10B29), anti-human CD4-PerCP (Clone# OKT4) and anti-human CD3-FITC (clone# SK7) were from Biolegend (San Diego, CA, USA); Indo-1-AM was from Molecular Probes (Eugene, OR).

Techniques: Purification, Incubation, Staining, Flow Cytometry, Comparison, Control

Journal: Frontiers in Veterinary Science

Article Title: Development of lateral flow assays to detect host proteins in cattle for improved diagnosis of bovine tuberculosis

doi: 10.3389/fvets.2023.1193332

Figure Lengend Snippet: Serial dilutions in triplicate for IFN-γ, IL-2, IL-6, CCL4, CXCL9, and CXCL10. Respective recombinant bovine cytokines were spiked in assay buffer and analyzed using UCP-LFAs. Shown are the ratios ( R ) obtained by dividing the signal at the respective test (T) lines by the signal at the flow control (FC) lines for each recombinant bovine protein. Mean values with error bars (±1 SD) are shown.

Article Snippet: The UCP conjugates for the six tested analytes were prepared to utilize the following antibodies: mouse anti-bovine IFN-γ (MT307; Mabtech, Stockholm, Sweden); mouse anti-bovine IL-2 (MT3B3; Mabtech, Stockholm, Sweden); chicken anti-bovine IL-6 (AHP2380B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL9 (AHP2369B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL10 (AHP2368B; Bio-Rad Laboratories, Hercules, CA, USA); and goat anti-bovine CCL4 (PBB0479B-050; Kingfisher Biotech, St Paul, MN, USA) at a concentration of 25 μg antibody per mg of UCP according to the methods described previously for the development of UCP-LFAs for the detection of M. leprae infection ( ).

Techniques: Recombinant, Control

Journal: Frontiers in Veterinary Science

Article Title: Development of lateral flow assays to detect host proteins in cattle for improved diagnosis of bovine tuberculosis

doi: 10.3389/fvets.2023.1193332

Figure Lengend Snippet: IFN-γ, IL-2, IL-6, CCL4, CXCL9, and CXCL10 levels in PPDb-stimulated whole blood supernatants were measured by UCP-LFAs. The ratios among groups were compared using the Kruskal–Wallis test with Dunn's multiple comparison post-test. N: Naive Animals ( n = 16, empty circles); V: BCG-vaccinated animals ( n = 10, green circles); V/C: BCG-vaccinated and Mycobacterium bovis challenged ( n = 12, blue circles); C: M. bovis challenged only ( n = 9, red circles). The bars of scatter dot plots show mean values, and error bars show ±1 SD. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The UCP conjugates for the six tested analytes were prepared to utilize the following antibodies: mouse anti-bovine IFN-γ (MT307; Mabtech, Stockholm, Sweden); mouse anti-bovine IL-2 (MT3B3; Mabtech, Stockholm, Sweden); chicken anti-bovine IL-6 (AHP2380B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL9 (AHP2369B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL10 (AHP2368B; Bio-Rad Laboratories, Hercules, CA, USA); and goat anti-bovine CCL4 (PBB0479B-050; Kingfisher Biotech, St Paul, MN, USA) at a concentration of 25 μg antibody per mg of UCP according to the methods described previously for the development of UCP-LFAs for the detection of M. leprae infection ( ).

Techniques: Comparison

Journal: Frontiers in Veterinary Science

Article Title: Development of lateral flow assays to detect host proteins in cattle for improved diagnosis of bovine tuberculosis

doi: 10.3389/fvets.2023.1193332

Figure Lengend Snippet: Receiver operator curve (ROC) analysis.

Article Snippet: The UCP conjugates for the six tested analytes were prepared to utilize the following antibodies: mouse anti-bovine IFN-γ (MT307; Mabtech, Stockholm, Sweden); mouse anti-bovine IL-2 (MT3B3; Mabtech, Stockholm, Sweden); chicken anti-bovine IL-6 (AHP2380B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL9 (AHP2369B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL10 (AHP2368B; Bio-Rad Laboratories, Hercules, CA, USA); and goat anti-bovine CCL4 (PBB0479B-050; Kingfisher Biotech, St Paul, MN, USA) at a concentration of 25 μg antibody per mg of UCP according to the methods described previously for the development of UCP-LFAs for the detection of M. leprae infection ( ).

Techniques: Significance Assay

Journal: Frontiers in Veterinary Science

Article Title: Development of lateral flow assays to detect host proteins in cattle for improved diagnosis of bovine tuberculosis

doi: 10.3389/fvets.2023.1193332

Figure Lengend Snippet: ROC curve analysis of IFN-γ (red), IL-2 (green), IL-6 (pink), CCL4 (yellow), CXCL9 (black), and CXCL10 (blue) detected in PPDb-stimulated whole blood supernatants to discriminate naïve (N) from Mycobacterium bovis -challenged group (C) (A) , BCG-vaccinated (V) from M. bovis -challenged group (C) (B) , and BCG-vaccinated group (V) from those that were M. bovis -challenged post-BCG (V/C) (C) .

Article Snippet: The UCP conjugates for the six tested analytes were prepared to utilize the following antibodies: mouse anti-bovine IFN-γ (MT307; Mabtech, Stockholm, Sweden); mouse anti-bovine IL-2 (MT3B3; Mabtech, Stockholm, Sweden); chicken anti-bovine IL-6 (AHP2380B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL9 (AHP2369B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL10 (AHP2368B; Bio-Rad Laboratories, Hercules, CA, USA); and goat anti-bovine CCL4 (PBB0479B-050; Kingfisher Biotech, St Paul, MN, USA) at a concentration of 25 μg antibody per mg of UCP according to the methods described previously for the development of UCP-LFAs for the detection of M. leprae infection ( ).

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Development of lateral flow assays to detect host proteins in cattle for improved diagnosis of bovine tuberculosis

doi: 10.3389/fvets.2023.1193332

Figure Lengend Snippet: A NUM score was calculated based on IFN-γ, IL-2, IL-6, CCL4, CXCL9, and CXCL10 ratios in PPDb-stimulated whole blood supernatants for naïve (N; n = 16), BCG-vaccinated (V; n = 10), and Mycobacterium bovis -challenged animals with (V/C; n = 12) or without prior BCG vaccination (C; n = 9). The NUM score ( y -axis) combines the results of six proteins, indicating the number of proteins with levels above a threshold based on the maximal Youden's index for each marker . Group differences were determined using the Kruskal-Wallis test; the statistical significance level used was *** p < 0.001; **** p < 0.0001 (A) . Heatmap showing Pearson correlation among the ratios of the evaluated host proteins. The color corresponds to the Pearson R value as indicated in each square (B) .

Article Snippet: The UCP conjugates for the six tested analytes were prepared to utilize the following antibodies: mouse anti-bovine IFN-γ (MT307; Mabtech, Stockholm, Sweden); mouse anti-bovine IL-2 (MT3B3; Mabtech, Stockholm, Sweden); chicken anti-bovine IL-6 (AHP2380B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL9 (AHP2369B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL10 (AHP2368B; Bio-Rad Laboratories, Hercules, CA, USA); and goat anti-bovine CCL4 (PBB0479B-050; Kingfisher Biotech, St Paul, MN, USA) at a concentration of 25 μg antibody per mg of UCP according to the methods described previously for the development of UCP-LFAs for the detection of M. leprae infection ( ).

Techniques: Marker

Journal: Frontiers in Veterinary Science

Article Title: Development of lateral flow assays to detect host proteins in cattle for improved diagnosis of bovine tuberculosis

doi: 10.3389/fvets.2023.1193332

Figure Lengend Snippet: NUM scores for biomarker signature.

Article Snippet: The UCP conjugates for the six tested analytes were prepared to utilize the following antibodies: mouse anti-bovine IFN-γ (MT307; Mabtech, Stockholm, Sweden); mouse anti-bovine IL-2 (MT3B3; Mabtech, Stockholm, Sweden); chicken anti-bovine IL-6 (AHP2380B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL9 (AHP2369B; Bio-Rad Laboratories, Hercules, CA, USA); rabbit anti-bovine CXCL10 (AHP2368B; Bio-Rad Laboratories, Hercules, CA, USA); and goat anti-bovine CCL4 (PBB0479B-050; Kingfisher Biotech, St Paul, MN, USA) at a concentration of 25 μg antibody per mg of UCP according to the methods described previously for the development of UCP-LFAs for the detection of M. leprae infection ( ).

Techniques: Biomarker Discovery