Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of IL-6-IL-27 Complex in Host Antiviral Immune Response.

doi: 10.4049/jimmunol.2100179

Figure Lengend Snippet: FIGURE 1. Interactions between the subunits of IL-6 and IL-27 in vivo and in vitro. (AC) CoIP and immunoblotting of serum samples were performed with the indicated Abs. IL-6 interacted with IL-27A (A), IL-6 interacted with EBi3 (B), and IL-27A interacted with EBi3 (C). (DG) HEK293T cells were transfected with the indicated plasmids for 24 h. CoIP and immunoblotting were performed with the indicated Abs. IL-6-3HA interacted with IL-27A-V5 (D), IL-6-3HA interacted with EBi3-Myc (E), IL-27A-V5 interacted with EBi3-Myc (F), and IL-6-3HA interacted with IL-27-V5 (G). (H) Colocalization of IL-6 and IL-27 was detected by confocal microscopy. A549 cells were transfected with IL-6-eGFP-KDEL and IL-27-DsRed-KDEL for 24 h, and the nuclei were stained with DAPI before observation using confocal microscopy. All experiments were repeated at least three times with consistent results.

Article Snippet: Abs against V5 tag (66007-1-Ig), EBi3 (12371-1-AP), myxovirus resistance A (MxA; 13750-1-AP), MAVS (14341-1-AP), TGFb-activated kinase 1 (12330-2-AP), IRF3 (11312-1-AP), NF-kB p65 (10745- 1-AP), NF-kB p50 (14220-1-AP), Lamin A/C (10298-1-AP), GAPDH (60004-1-Ig), and b-actin (60008-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: In Vivo, In Vitro, Western Blot, Transfection, Confocal Microscopy, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of IL-6-IL-27 Complex in Host Antiviral Immune Response.

doi: 10.4049/jimmunol.2100179

Figure Lengend Snippet: FIGURE 2. Construction of the IL-6IL-27 complex and its antiviral activity. (A) A schematic of the IL-6IL-27 construction via flexible self-cleaved linker is shown. Immunoblotting analysis of IL-6, EBi3, IL-27A, and b-actin of HEK293T cells transfected with empty vector pcDNA3.1(1) or IL-6IL-27 complex expression plasmids for 24 h. (B) A549 cells were transfected with indicated plasmids for 24 h followed by H1N1 infection (MOI = 1) for 12 h. The NP-specific mRNA, cRNA, and vRNA levels were measured using QRT-PCR. (C) RD cells were transfected with the indicated plasmids for 24 h fol- lowed by EV71 infection (MOI = 1) for 24 h. The VP1 mRNA levels were measured using QRT-PCR. (D) Vero cells were (Figure legend continues)

Article Snippet: Abs against V5 tag (66007-1-Ig), EBi3 (12371-1-AP), myxovirus resistance A (MxA; 13750-1-AP), MAVS (14341-1-AP), TGFb-activated kinase 1 (12330-2-AP), IRF3 (11312-1-AP), NF-kB p65 (10745- 1-AP), NF-kB p50 (14220-1-AP), Lamin A/C (10298-1-AP), GAPDH (60004-1-Ig), and b-actin (60008-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: Activity Assay, Western Blot, Transfection, Plasmid Preparation, Expressing, Infection, Quantitative RT-PCR

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of IL-6-IL-27 Complex in Host Antiviral Immune Response.

doi: 10.4049/jimmunol.2100179

Figure Lengend Snippet: FIGURE 5. The IL-6IL-27 complex interacts with MAVS. (AC) A549 cells were transfected with increasing amounts of the indicated expression plas- mids (wedge; 100, 200, and 300 ng) of the IL-6IL-27 complex expression plasmid and luciferase reporter plasmids containing the IFN-b (A), NF-kB (B), and ISRE (C) promoter for 24 h followed by VSV infection (MOI = 1) for 24 h. Reporter assays were performed. pRL-TK was used as an internal control. (DI) HEK293T cells were transfected with the indicated plasmids for 24 h. CoIP and immunoblotting were performed with the indicated Abs. IL-6-3HA interacted with Flag-MAVS (D), IL-27A-V5 interacted with Flag-MAVS (E), EBi3-Myc interacted with Flag-MAVS (F), IL-6-HA interacted with Flag- TBK1 (G), IL-27A-V5 interacted with Flag-TBK1 (H), and EBi3-Myc interacted with Flag-TBK1 (I). All experiments were repeated at least three times with consistent results.

Article Snippet: Abs against V5 tag (66007-1-Ig), EBi3 (12371-1-AP), myxovirus resistance A (MxA; 13750-1-AP), MAVS (14341-1-AP), TGFb-activated kinase 1 (12330-2-AP), IRF3 (11312-1-AP), NF-kB p65 (10745- 1-AP), NF-kB p50 (14220-1-AP), Lamin A/C (10298-1-AP), GAPDH (60004-1-Ig), and b-actin (60008-1-Ig) were purchased from ProteinTech Group (Wuhan, China).

Techniques: Transfection, Expressing, Plasmid Preparation, Luciferase, Infection, Control, Western Blot

Journal: Mediators of Inflammation

Article Title: Associations of Trauma Severity with Mean Platelet Volume and Levels of Systemic Inflammatory Markers (IL1 β , IL6, TNF α , and CRP)

doi: 10.1155/2016/9894716

Figure Lengend Snippet: Comparison of biochemical markers within between study groups and controls.

Article Snippet: Serum TNF α , IL1 β , and IL6 levels were measured using Boster ELISA kits (Freemont, CA, USA) and Bio-Tek (Winooski, VT, USA) ELx50 and ELx800 devices.

Techniques: Comparison, Control

Journal: BMC Biotechnology

Article Title: Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample

doi: 10.1186/1472-6750-14-63

Figure Lengend Snippet: Curve fitting for IL-1β and IP-10. Visualization of the regression line (blue) fitted for the theoretical concentrations and membrane’s fluorescence readouts of IL-1β (a) and IP-10 (b) . The dots are compactly distributed around the predicted line, and the unexplained error is small. The red lines delimited the 95% of CI. The equations of the curves are: a) y = 561.73 × + 8.97 with Pearson’s r = 0.97; b) y = 1830.6 × – 79.4 with Pearson’s r = 0.91. The red dashed lines delimited the 95% confidence interval.

Article Snippet: Recombinant Human IL-1 beta and Recombinant Human CXCL10/IP-10 from R& D Systems (R& D Systems™) were reconstituted according to the manufacturer’s instructions.

Techniques: Fluorescence

Journal: BMC Biotechnology

Article Title: Improving membrane based multiplex immunoassays for semi-quantitative detection of multiple cytokines in a single sample

doi: 10.1186/1472-6750-14-63

Figure Lengend Snippet: Model diagnostics of the curve fitting of IL-1β (a) and IP-10 (b). "Residuals vs. Fitted" and "Scale-Location" show that a trend appears in the residuals distribution for concentrations below 400 ng/mL. The “Q-Q plot” suggests that residuals are not normally distributed. Whilst this does not affect the goodness of the coefficient estimation, it impairs the t-distribution and thus renders the use of p-values meaningless. The Cook’s distance suggests that 2 of the most diluted samples could be considered outliers in their influence on the model fitting. However, Cook’s distance has no unambiguous interpretation and the best decision is to look at the original distribution, where the 2 incriminated points show no indication of anomalous position. The red dashed lines delimited the 95% of CI.

Article Snippet: Recombinant Human IL-1 beta and Recombinant Human CXCL10/IP-10 from R& D Systems (R& D Systems™) were reconstituted according to the manufacturer’s instructions.

Techniques:

Journal: Mediators of Inflammation

Article Title: DAMPs Synergize with Cytokines or Fibronectin Fragment on Inducing Chondrolysis but Lose Effect When Acting Alone

doi: 10.1155/2017/2642549

Figure Lengend Snippet: In the initial two experiments with bovine chondrocytes, HMGB1 synergized with IL-1 β on upregulating metalloproteinase production while MTDs (fMLF and CpG DNA) or HMGB1 alone showed little effect. Cells were harvested from cartilage in bovine stifle joints and cultured for 1 week. Passage 1 cells were treated with MTDs, HMGB1, and/or IL-1 β . After 24 hrs, culture medium was collected, dialyzed, and concentrated. The expression of MMPs in each medium sample was determined with Western blotting. U. C. = untreated control; CpG = CpG-rich DNA; CpG N. C. = CpG-rich DNA negative control.

Article Snippet: This could be attributed to the fact that the recombinant IL-1 β protein used in our study was human origin (R&D Systems, Cat # 201-LB) and might cause less strength of stimulation to bovine cells than to human cells.

Techniques: Cell Culture, Expressing, Western Blot, Control, Negative Control

Journal: Mediators of Inflammation

Article Title: DAMPs Synergize with Cytokines or Fibronectin Fragment on Inducing Chondrolysis but Lose Effect When Acting Alone

doi: 10.1155/2017/2642549

Figure Lengend Snippet: Observations made in bovine chondrocytes were further verified in the same type of cells in humans. Articular chondrocytes were isolated from the ankle cartilage of an amputated patient. Passage 2 cells were treated with DAMPs and/or Fn-f (a) or IL-1 β (b) for 24 hrs. Medium samples were analyzed for MMP-3 expression with Western blotting. Relative intensity of protein bands on each blot was measured and plotted, respectively.

Article Snippet: This could be attributed to the fact that the recombinant IL-1 β protein used in our study was human origin (R&D Systems, Cat # 201-LB) and might cause less strength of stimulation to bovine cells than to human cells.

Techniques: Isolation, Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: DAMPs Synergize with Cytokines or Fibronectin Fragment on Inducing Chondrolysis but Lose Effect When Acting Alone

doi: 10.1155/2017/2642549

Figure Lengend Snippet: Observations made in bovine chondrocytes were further verified in the same type of cells in humans. Articular chondrocytes were isolated from ankle cartilage of an amputated patient. Passage 2 cells were treated with DAMPs and/or Fn-f (a) or IL-1 β (b) for 24 hrs. Medium samples were analyzed for expression of MMP-13 and ADAMTS-5 with Western blotting. Furthermore, the induction pattern of ADAM-8, a newly discovered fibronectinase, was examined with the same technique.

Article Snippet: This could be attributed to the fact that the recombinant IL-1 β protein used in our study was human origin (R&D Systems, Cat # 201-LB) and might cause less strength of stimulation to bovine cells than to human cells.

Techniques: Isolation, Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: DAMPs Synergize with Cytokines or Fibronectin Fragment on Inducing Chondrolysis but Lose Effect When Acting Alone

doi: 10.1155/2017/2642549

Figure Lengend Snippet: Protein expression of iNOS, an inflammation-pathway-downstream effector, was only induced by IL-1 β or TNF- α not by DAMPs. Bovine (a) or human (b) articular chondrocytes were treated with DAMPs or cytokines or Fn-f or in combination for 24 hrs. Cells were then lyzed, and the lysates were examined for expression of iNOS with Western blotting.

Article Snippet: This could be attributed to the fact that the recombinant IL-1 β protein used in our study was human origin (R&D Systems, Cat # 201-LB) and might cause less strength of stimulation to bovine cells than to human cells.

Techniques: Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: DAMPs Synergize with Cytokines or Fibronectin Fragment on Inducing Chondrolysis but Lose Effect When Acting Alone

doi: 10.1155/2017/2642549

Figure Lengend Snippet: The synergism between HMGB1 and IL-1 β was replicable in the 3rd experiment with bovine chondrocytes. Such synergism was also observed between DAMPs and TNF- α but to a lesser extent. However, DAMPs were unable to synergize with Fn-f on the upregulation of MMPs while the synergism between cytokines and Fn-f was observed. Moreover, DAMPs alone or in combination did not evoke secretion of MMPs from bovine chondrocytes, which was consistent with what was observed in previous two experiments. Passage 1 bovine chondrocytes were treated with DAMPs with or without Fn-f (a), with or without proinflammatory cytokines (TNF- α or IL-1 β ) (b) for 24 hrs. Culture medium was then examined for MMP expression with Western blotting. In addition, some cells were insulted with culture medium containing soluble substances released from injured cartilage which was bluntly impacted at 7 or 14 J/cm 2 and cultured for 1 day. After 24 hrs of incubation, culture medium was resolved by SDS-PAGE side by side with medium samples from cultures treated with HMGB1, or MTDs, or Fn-f, or TNF- α . Expression of MMP-3 was then examined with immunoblotting (c). U. C. = untreated control; U. I. = unimpacted control; I. = impacted.

Article Snippet: This could be attributed to the fact that the recombinant IL-1 β protein used in our study was human origin (R&D Systems, Cat # 201-LB) and might cause less strength of stimulation to bovine cells than to human cells.

Techniques: Expressing, Western Blot, Cell Culture, Incubation, SDS Page, Control

Journal: Mediators of Inflammation

Article Title: DAMPs Synergize with Cytokines or Fibronectin Fragment on Inducing Chondrolysis but Lose Effect When Acting Alone

doi: 10.1155/2017/2642549

Figure Lengend Snippet: Summary of various types of treatment involving DAPMs examined in the study.

Article Snippet: This could be attributed to the fact that the recombinant IL-1 β protein used in our study was human origin (R&D Systems, Cat # 201-LB) and might cause less strength of stimulation to bovine cells than to human cells.

Techniques: Expressing

Journal: American Journal of Cancer Research

Article Title: Different races have different immune microenvironments: comparison of White and Asian patients with liver cancer

doi:

Figure Lengend Snippet: Kaplan-Meier plots for Asian HCC patients based on the expression levels of CLEC5A (A) and GPM6A (B). Kaplan-Meier plots for White HCC patients based on the expression levels of IL-18RAP (C) and SIM1 (D).

Article Snippet: Colony formation assay Cells (1×10 3 ) treated with recombinant IL-18 protein, or IL-18RAP protein, or both (Sino Biological, Beijing, China) were plated in 24-well plates.

Techniques: Expressing

Journal: American Journal of Cancer Research

Article Title: Different races have different immune microenvironments: comparison of White and Asian patients with liver cancer

doi:

Figure Lengend Snippet: IL-18RAP (A) and IL-18 (B) serum levels from the HCC patients and healthy controls were detected using ELISA. Kaplan-Meier curves of the survival rate of the HCC patients based on their IL-18RAP (C) and IL-18 (D) serum levels. Correlation between IL-18 and IL-18RAP in healthy volunteers (E) and HCC patients (F).

Article Snippet: Colony formation assay Cells (1×10 3 ) treated with recombinant IL-18 protein, or IL-18RAP protein, or both (Sino Biological, Beijing, China) were plated in 24-well plates.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: American Journal of Cancer Research

Article Title: Different races have different immune microenvironments: comparison of White and Asian patients with liver cancer

doi:

Figure Lengend Snippet: Serum IL-18RAP and IL-18 associated with the clinicopathological features of 76 hepatocellular carcinoma patients

Article Snippet: Colony formation assay Cells (1×10 3 ) treated with recombinant IL-18 protein, or IL-18RAP protein, or both (Sino Biological, Beijing, China) were plated in 24-well plates.

Techniques: Infection

Journal: American Journal of Cancer Research

Article Title: Different races have different immune microenvironments: comparison of White and Asian patients with liver cancer

doi:

Figure Lengend Snippet: The functions of IL-18RAP and GPM6A in HCC cells. A. The interaction between IL-18 and IL-18RAP was simulated using HEX 8.0.0 software. B. The proliferation ratio was analyzed using colony formation assay. C. The proportion of apoptotic cells was determined by Annexin-V/FITC and PI double staining. D. Western blot analysis of the Smad signaling pathway. GAPDH was used as an internal loading control.

Article Snippet: Colony formation assay Cells (1×10 3 ) treated with recombinant IL-18 protein, or IL-18RAP protein, or both (Sino Biological, Beijing, China) were plated in 24-well plates.

Techniques: Software, Colony Assay, Double Staining, Western Blot