Journal: iScience

Article Title: Macrophages treated with interferons induce different responses in lymphocytes via extracellular vesicles

doi: 10.1016/j.isci.2024.109960

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti CD261/TRAIL-R1 (clone S35-934) Conjugated BV786 , BD OptiBuild , Cat#744711.

Techniques: Recombinant, Protein Extraction, Protease Inhibitor, Pore Size, Electrophoresis, Staining, Software, Cytometry

Journal: Cell Reports Medicine

Article Title: Lymph node and tumor-associated PD-L1 + macrophages antagonize dendritic cell vaccines by suppressing CD8 + T cells

doi: 10.1016/j.xcrm.2023.101377

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Article Snippet: Pe/cyanine7 anti-mouse TRAIL (clone N2B2) , Biolegend , Cat#109311 RRID: AB_2721675.

Techniques: Recombinant, Lysis, Protease Inhibitor, Western Blot, Staining, Stripping, Liposomes, CRISPR, MTS Assay, ATP Assay, Cell Isolation, Enzyme-linked Immunosorbent Assay, Conjugation Assay, Selection, Vaccines, Single-cell Analysis, RNA Sequencing Assay, Mutagenesis, Microarray, Purification, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: HCMV-secreted glycoprotein gpUL4 inhibits TRAIL-mediated apoptosis and NK cell activation

doi: 10.1073/pnas.2309077120

Figure Lengend Snippet: UL4 binds and inhibits NK cells in a TRAIL-dependent manner. ( A ) PBMCs were stimulated overnight with IFNα, or unstimulated, then stained for CD3/CD56 along with either TRAIL ( A ) or gpUL4-V5 ( B ), before gating on the CD3 − CD56 + population, and plotting the level of TRAIL or UL4 staining. ( C ) Mock or HCMV-infected HFFF-hTert were incubated with PBMC for 5 h in the presence of the indicated proteins (5 μg/mL); then, the level of degranulating NK cells (CD3 − CD56 + ) was assessed. ( D ) NK cells and HFFF-hTert were mixed in the presence of gpUL4-V5 (5 μg/mL) protein. gpUL4-V5 was kept throughout the entire reaction (in Rxn), or PBMCs were pretreated with gpUL4-V5 and excess washed off (PBMC only), or HFFF-hTert were pretreated with protein and excess washed off (HFFF only), then the level of degranulating NK cells (CD3 − CD56 + ) assessed 5 h later. ( E ) A549-ACE2 targets infected with SARS-CoV-2 for 24 h were coincubated with NK cells for 5 h; then, the level of degranulating NK cells (CD3 − CD56 + ) was assessed. Two-way ANOVA with Tukey correction ( C and E ) and one-way ANOVA with Tukey correction ( D ): * P < 0.05, ** P < 0.01, *** P < 0.001, and *** P < 0.0001.

Article Snippet: After blocking, the plate was washed with PBS-T (PBS, 0.05% Tween 20) and incubated with 1 or 10 μg/mL of UL4 or TRAIL for 90 min at 37 °C followed by washes with PBS-T. Primary antibody in blocking buffer was added for 1 h at room temperature, washed with PBS-T and secondary antibody in blocking buffer was added for another 1 h at room temperature followed by washes with PBS-T. Primary antibodies used were mouse anti-V5 antibody (Bio-Rad) or mouse anti-TRAIL (cat.no ab10516, Abcam), and the secondary antibody was goat anti-mouse HRP (Santa Cruz).

Techniques: Staining, Infection, Incubation

Journal: iScience

Article Title: TRAIL-dependent apoptosis of peritoneal mesothelial cells by NK cells promotes ovarian cancer invasion

doi: 10.1016/j.isci.2023.108401

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Article Snippet: Mouse anti-human TRAIL-R2-PE , Biolegend , Cat# 307405; RRID: AB_314677.

Techniques: Recombinant, Polymer, Software

Journal: iScience

Article Title: TRAIL-dependent apoptosis of peritoneal mesothelial cells by NK cells promotes ovarian cancer invasion

doi: 10.1016/j.isci.2023.108401

Figure Lengend Snippet: TRAIL sensitivity of HPMC is linked to low expression of MHC class I and high expression of DR5 death receptor (A and B) Expression of MHC class I genes (HLA-A, HLA-B, HLA-C) (A) and TRAIL receptor genes (TNFRSF10A, TNFRSF10B, TNFRSF10C, TNFRSF10D). (B) In HPMC (from omentum) compared to tumor cells and TAT (from ascites) from HGSC patients according to RNA-Seq datasets as described previously. TPM values are depicted from n = 5 HPMC, n = 34 tumor cells and n = 6 TAT of different patients. (C) The expression of TRAIL receptors (DR4, DR5, DcR1, DcR2) on ex vivo tumor cells (n = 12 patients) and cultured HPMC from HGSC patients (n = 8 patients) and control HPMC originated from patients with benign gynecological diseases (Ctrl HPMC, n = 6 patients) was further validated on protein-level via flow cytometry. The geometric MFI minus the isotype control is depicted. (D) Overlay of representative histograms showing the TRAIL receptor expression (DR4, DR5, DcR1, DcR2) in comparison to the isotype controls for HPMC and tumor cells from HGSC patients are presented. The mean is shown by horizontal bars and vertical error bars represent the standard deviation. ∗ FDR < 0.05; ∗∗ FDR < 0.01; ∗∗∗ FDR < 0.001; ∗∗∗∗ FDR < 0.0001 determined by unpaired t test and Benjamini-Hochberg adjustment (ns: not significant).

Article Snippet: Mouse anti-human TRAIL-R1-APC , Biolegend , Cat# 307207; RRID: AB_2256112.

Techniques: Expressing, RNA Sequencing Assay, Ex Vivo, Cell Culture, Flow Cytometry, Comparison, Standard Deviation

Journal: iScience

Article Title: TRAIL-dependent apoptosis of peritoneal mesothelial cells by NK cells promotes ovarian cancer invasion

doi: 10.1016/j.isci.2023.108401

Figure Lengend Snippet:

Article Snippet: Mouse anti-human TRAIL-R1-APC , Biolegend , Cat# 307207; RRID: AB_2256112.

Techniques: Recombinant, Polymer, Software