Journal: The Journal of Immunology

Article Title: Response to Comment on “Decoy Receptor 3 Expression in AsPC-1 Human Pancreatic Adenocarcinoma Cells via the Phosphatidylinositol 3-Kinase-, Akt-, and NF-κB-Dependent Pathway”

doi: 10.4049/jimmunol.0990054

Figure Lengend Snippet: FIGURE 1. ELISA and immunoprecipitation for determination of DcR3 protein expression in the human pancreatic cancer cell lines AsPC-1 and PANC-1. A, Cells (1 105) were cultured in 24-well plates, the supernatants were collected after 48 h of incubation, DcR3 levels were detected by ELISA, and the cells were collected to quantify that the protein concentration was equal. B, Equal amounts of concentrated medium from cells was immunopre- cipitated (IP) with 1 g of anti-DcR3 Ab, followed by Western blotting (WB) using the same DcR3 Ab. The GAPDH Ab was used for loading control. HT-29 cells were used as the negative control and SW480 cells were used as positive control for DcR3 expression, respectively. The blots shown are the representative of those obtained in three separate experiments. The data are the mean SEM for three separate experiments.

Article Snippet: We used a commercial DuoSet ELISA development system (catalog number DY142; R&D Systems) to detect DcR3 expression levels, and all of our results were comprehended within the detection range of this kit, which limited the analytic concentration below 12 ng/ml in accordance with the manufacturer’s suggestion.

Techniques: Enzyme-linked Immunosorbent Assay, Immunoprecipitation, Expressing, Cell Culture, Incubation, Protein Concentration, Western Blot, Control, Negative Control, Positive Control

Journal: Indian journal of pathology & microbiology

Article Title: A tissue microarray study of toll-like receptor 4, decoy receptor 3, and external signal regulated kinase 1/2 expressions in astrocytoma.

doi: 10.4103/0377-4929.188122

Figure Lengend Snippet: Figure 1: H and E staining of (A0) normal brain, (B0) diffuse astrocytoma, (C0) anaplastic astrocytoma, and (D0) glioblastoma. Immunohistochemical analysis of toll-like receptor 4 in (A1) normal brain, (B1) diffuse astrocytoma, (C1) anaplastic astrocytoma, and (D1) glioblastoma; decoy receptor 3 in (A2) normal brain, (B2) diffuse astrocytoma, (C2) anaplastic astrocytoma, and (D2) glioblastoma; and external signal regulated kinase in (A3) normal brain, (B3) diffuse astrocytoma, (C3) anaplastic astrocytoma, and (D3) glioblastoma (original magnification, ×400)

Article Snippet: Paraffin sections (5 μm thick) were dewaxed in xylene, rehydrated in an alcohol series, immersed in 3% hydrogen peroxide for 10 min to suppress endogenous peroxidase activity, heated (100°C) 30 min in 0.01 mol/L sodium citrate buffer (pH 6.0) to retrieve antigen, rinsed three times in phosphate buffered saline (PBS) for 5 min, and incubated 1 h at room temperature with mouse monoclonal antibody to human DcR3 (1:100 Santa Cruz Biotechnology, CA, USA), mouse monoclonal antibody to human TLR4 (1:100 Santa Cruz Biotechnology, CA, USA), and rabbit monoclonal anti‐human ERK1/2 antibody (1:100 Epitomics) diluted in PBS.

Techniques: Staining, Immunohistochemical staining

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: DcR3 is a potential novel diagnostic marker and therapeutic target in oral squamous cell carcinoma (OSCC) patients. Kaplan‐Meier curve compares the overall survival of cancer with high‐level or low‐level DcR3 protein products. Samples with immunohistochemistry (IHC) scores ≥4 were defined as having high DcR3 expression [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Diagnostic Assay, Marker, Immunohistochemistry, Expressing

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: Overexpression of DcR3 in oral cancer. A, Positive cytosol immunostaining of DcR3 in normal mucosa and oral cancer tissues. B, Quantitative polymerase chain reaction results from oral cancer tissues ( n = 30 patients) and their matched adjacent normal mucosal tissues ( n = 8 patients). C, DcR3 mRNA expression in human tongue cancer. Data were obtained from NCBI Gene Expression Omnibus profiles ( http://www.ncbi.nlm.nih.gov/geoprofiles ; Reporter: GDS4562). D, DcR3 protein in three tongue cancer cell lines was determined through Western blot analysis. Normal human gingival fibroblast (HGF) cells were used as negative control. P < 0.05 was considered statistically significant [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Over Expression, Immunostaining, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: Triptolide (TPL) inhibited tumor growth in DcR3‐overexpressing oral cancer patient‐derived tumor xenograft (PDTX) models. A, Changes in tumor volume in 134‐PDTX models ( n = 4) treated with TPL (0.15 mg/kg daily intraperitoneally) and phosphate‐buffered saline (PBS) (vehicle control; n = 4) for 28 days. Tumor diameters were measured twice weekly for 28 days using Vernier calipers; tumor volume was calculated and compared with those of controls. P < 0.05 was considered statistically significant. B, Tumor mass was weighed after the mice were sacrificed. C, No significant change was observed in the body weight of the mice compared with that of the vehicle controls. D, Hematoxylin and eosin staining and immunohistochemistry were performed after administration of TPL or PBS (vehicle control). The 134‐PDTX model stained positive for DcR3. E, The SAS xenograft model stained positive for DcR3. Immunodetectable proteins were stained brown; nuclei were counterstained blue. Original magnification: ×400 [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Derivative Assay, Staining, Immunohistochemistry

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: Triptolide (TPL) decreased DcR3 expression in oral cancer. A, Assessment of cell proliferation and viability using the methylene blue assay in the three oral cancer cell lines treated with varying concentrations of TPL (0‐100 nM) or DMSO (1 μL/mL) for 24 and 48 hours. B, Western blot analysis for DcR3 after SAS cells were treated with TPL for 24 and 48 hours

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Expressing, Western Blot

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: Triptolide (TPL) repressed MTA1 expression in oral cancer. A, Correlation analysis of DcR3 and MTA1 expression in oral squamous cell carcinoma (OSCC) tissue microarray. B, Hematoxylin and eosin staining and immunohistochemistry were performed after administration of TPL or phosphate‐buffered saline (PBS) (vehicle control). The 134‐PDTX and SAS xenograft models stained positive for MTA1. Immunodetectable proteins were stained brown; nuclei were counterstained blue. Original magnification: ×400. C, MTA1 protein in 3 tongue cancer cell lines was determined through Western blot analysis. Normal human gingival fibroblast (HGF) cells were used as negative control. D, Western blot analysis for MTA1 after SAS cells were treated with TPL for 24 and 48 hours [Color figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Expressing, Microarray, Staining, Immunohistochemistry, Western Blot, Negative Control

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: MTA1 regulated DcR3 expression in SAS cancer cells. A, DcR3 expression decreased after introduction of shMTA1 in SAS cells. B, DcR3 was overexpressed after MTA1 was overexpressed, and was subsequently downregulated through triptolide (TPL) treatment in SAS cells. C, Addition of the DcR3‐overexpressed vector was not associated with changes in MTA1 expression in SAS cells

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Expressing, Plasmid Preparation

Journal: Head & Neck

Article Title: Anti‐oral cancer effects of triptolide by downregulation of DcR3 in vitro, in vivo, and in preclinical patient‐derived tumor xenograft model

doi: 10.1002/hed.25554

Figure Lengend Snippet: Associated between DcR3 expression and multiple clinicopathological parameters in oral squamous cell carcinoma (OSCC)

Article Snippet: The plasmids expressing MTA1 and DcR3 were obtained from OriGene (Rockville, Maryland).

Techniques: Expressing