Journal: Journal of leukocyte biology

Article Title: Activated NK cells kill hepatic stellate cells via p38/PI3K signaling in a TRAIL-involved degranulation manner.

doi: 10.1002/JLB.2A0118-031RR

Figure Lengend Snippet: FIGURE 5 IL-18/poly I:C-primed NK cells kill hepatic stellate cells in a TRAIL- involved degranulation manner. IL-18 and poly I:C-primed NK cells were treated by anti-TRAIL or isotype antibody before coculture. HSC death was measured (A), and NK cell degranulation was evaluated by CD107a expression (C). Soluble TRAIL was added to cell coculture, the cell death of HSCs (B) and CD107a expression of NK cells (D) was shown. Results were shown as mean ± SEM of 6 independent experiments performed with 6 different donor NK cells. *P < 0.05, **P < 0.01, ***P < 0.001, paired t-test. The kinetics of the activated NK cell-mediated LX2 cell apoptosis were observed by using live-cell imaging (E).

Article Snippet: Soluble TRAIL (R&D System; Catalog number: 375-TL-010) was added to HSCs half hour earlier than NK cells in coculture.

Techniques: Expressing, Live Cell Imaging

Journal: Journal of leukocyte biology

Article Title: Activated NK cells kill hepatic stellate cells via p38/PI3K signaling in a TRAIL-involved degranulation manner.

doi: 10.1002/JLB.2A0118-031RR

Figure Lengend Snippet: FIGURE 6 IL-18/poly I:C-primed hepatic NK cells kill HSCs in a TRAIL-involved degranulation manner. Liver NK cells were purified from speci- men, then primed by IL-18 and/or poly I:C before cocultured with primary HSCs as above. CD107a expression of liver NK cells (A) and PI/AnnexinV level of primary HSCs (B) were detected. Anti-TRAIL or isotype antibody were used to treat IL-18/poly I:C-primed liver NK cells before cocultured with primary HSCs, then CD107a expression of liver NK cells (C) and the cell death of primary HSCs (D) was measured. Results were shown as mean ± SEM of 5 independent experiments performed with 5 different donor NK cells. *P < 0.05, **P < 0.01, paired t-test.

Article Snippet: Soluble TRAIL (R&D System; Catalog number: 375-TL-010) was added to HSCs half hour earlier than NK cells in coculture.

Techniques: Purification, Expressing

Journal: Arthritis and rheumatism

Article Title: Mechanisms and clinical relevance of TRAIL-triggered responses in the synovial fibroblasts of patients with rheumatoid arthritis.

doi: 10.1002/art.30181

Figure Lengend Snippet: Figure 4. Rheumatoid arthritis fibroblast-like synoviocytes (RA FLS) produce low levels of the inflammatory cytokines interleukin-6 (IL-6) and IL-8 when stimulated with TRAIL. RA FLS (n 5) were treated with TRAIL (1 nM) or tumor necrosis factor (TNF; 0.5 nM) for 72 hours, and supernatants were analyzed for IL-6 (A), IL-8 (B), and RANTES (C) secretion by enzyme-linked immunosorbent assay. Cy- tokine levels were analyzed using Wilcoxon’s test. Values are the mean SEM. NS not stimulated.

Article Snippet: Serum levels of TRAIL and OPG were measured using a commercially available enzyme-linked immunosorbent assay (ELISA) (Quantikine human OPG ELISA Kit [R&D Systems] and a human TRAIL ELISA [Diaclone]).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Molecular cancer research : MCR

Article Title: A genome-wide loss-of-function screen identifies SLC26A2 as a novel mediator of TRAIL resistance

doi: 10.1158/1541-7786.MCR-16-0234

Figure Lengend Snippet: (A) Flow cytometry analysis in BJAB-LEXR cells with stable, lentiviral, shRNAs against SLC26A2 or a Non-targeting shRNA after 24hrs of treatment with TRAIL (20ng/mL) to assess propidium iodide (PI) positive cells as an indication of dead or late apoptotic cells. Representative experiment is shown (of 2 experiments). (B) MDA-231 SENS and TRAILR cells plated in replicates of 6 were treated with TRAIL (100ng/mL) and a green fluoregenic substrate for activated-caspase3/7. Green fluorescence was measured every 2 hours with 100ng/mL TRAIL by light microscopy using real time in-vitro imaging. Statistical significance assessed by 2-way ANOVA with a Tukey post-test. A representative figure is shown (of 2 experiments). (C) Triplicate samples of 231 SEN and 231 TRAILR were analyzed for RNA expression of SLC26A2, DR4, and DR5, using microarray analysis. The expression is presented as a heat map relative to an average expression of a combination of constitutive housekeeping genes. RNA and protein were isolated from 231 SEN cells and 231 TRAILR cells as well as 231 TRAILR cells transduced with non-targeting control shRNA (non-coding) or with either of 2 shRNA targeting SLC26A2 (SLC26A2 sh1 and SLC26A2 sh2, respectively). mRNA expression of (D) SLC26A2, (E) DR4 and (F) DR5 was determined by real time quantitative PCR. The expression is relative to the expression of GAPDH and was normalized to non-targeting shRNA vector-expressing cells. Representative experiments are shown (of ≥ 3 experiments) with each experiment performed in triplicate. Statistical significance assessed by 1-way ANOVA. Protein expression of these genes as well as β-actin was analyzed by Western blot as shown in (G). Surface expression of DR4 and DR5 was analyzed by flow cytometry as depicted in (H) and (I), respectively. Representative experiments are shown (of ≥ 3 experiments). Statistical significance assessed by 1-way ANOVA.

Article Snippet: 10μl/1X10 6 of PE conjugated antibodies against DR4 (FAB347P, R&D Systems) and DR5 (FAB6311P, R&D Systems) were then added and allowed to incubate for 30 minutes at room temperature in the dark followed by 3 washes with Flow Cytometry Staining Buffer.

Techniques: Flow Cytometry, shRNA, Fluorescence, Light Microscopy, In Vitro, Imaging, RNA Expression, Microarray, Expressing, Isolation, Transduction, Real-time Polymerase Chain Reaction, Plasmid Preparation, Western Blot