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 FasL-PE , Miltenyi Biotec , Cat# 130-118-491; RRID: AB_2751526.

Techniques: Control, Recombinant, Polymer, Software

Journal: Cell Death & Disease

Article Title: ZNRF1 deficiency disrupts Fas ligand trafficking and immune balance

doi: 10.1038/s41419-026-08566-8

Figure Lengend Snippet: a Immunoblot analysis of FasL and ZNRF1 in control ( sgCtrl ) and Znrf1 -targeted RAW264.7 macrophage clones ( sgZnrf1 #1 and #2) stimulated with lipopolysaccharide (LPS, 100 ng/mL) for the indicated times (0, 16, and 24 h). β-Actin served as a loading control. Numbers beneath the ZNRF1 blot indicate densitometric quantification after lane-specific background subtraction (ZNRF1/β-actin), normalized to sgCtrl 0 h (= 1). b Representative flow-cytometry histograms of surface FasL in sgCtrl and sgZnrf1 macrophages under vehicle control conditions or after LPS stimulation for 16 or 24 h. c Quantification of surface FasL mean fluorescence intensity (MFI) from ( b ) ( n = 3 biological replicates per group; representative of three independent experiments). d Immunoblot analysis of FasL and ZNRF1 in bone-marrow-derived macrophages (BMDMs) from Znrf1 f/f and Znrf1 Δ mice stimulated with LPS (100 ng/mL) for the indicated times. e Representative flow-cytometry histograms of surface FasL in Znrf1 f/f and Znrf1 Δ BMDMs following LPS stimulation. f Quantification of surface FasL MFI from ( e ) ( n = 3 biological replicates per group; representative of two independent experiments). g Representative flow-cytometry histograms of surface FasL in Znrf1 f/f and Znrf1 Δ BMDMs stimulated with live E. coli at the indicated multiplicities of infection (MOI). h Quantification of surface FasL MFI from ( g ) ( n = 3 biological replicates per group; representative of two independent experiments). Data are shown as mean ± SD. * p < 0.05, ** p < 0.01; n.s., not significant (unpaired two-tailed t-test).

Article Snippet: FasL cDNA, obtained from Origene (Rockville, MD), was polymerase chain reaction–amplified and cloned into a C-terminally FLAG-tagged pLKO-AS2 lentiviral vector.

Techniques: Western Blot, Control, Clone Assay, Flow Cytometry, Fluorescence, Derivative Assay, Infection, Two Tailed Test

Journal: Cell Death & Disease

Article Title: ZNRF1 deficiency disrupts Fas ligand trafficking and immune balance

doi: 10.1038/s41419-026-08566-8

Figure Lengend Snippet: a Co-immunoprecipitation of Munc18-2 and Syntaxin-3 in control ( sgControl ) and Znrf1 -deficient ( sgZnrf1 , two sgRNAs) murine macrophages after LPS (100 ng/mL, 24 h) stimulation. Cell lysates were immunoprecipitated with anti-Munc18-2 or IgG (control), followed by immunoblot analysis with specified antibodies. b Confocal microscopy of murine macrophages stained for FasL (green), Munc18-2 (blue), and Syntaxin-3 (red) under vehicle or LPS (20 h) conditions. Right panels: Signal profiles along the indicated dashed lines in each cell, showing fluorescence intensity for FasL (green), Munc18-2 (blue), and Syntaxin 3 (red). Scale bars, 5 µm. c In RAW264.7 macrophages, knockdown of Munc18-2 ( Stxbp2 ) and Stx3 ( Syntaxin 3 ) was achieved using siRNA targeting two distinct sequences. Following this genetic manipulation, the cells were harvested and analyzed for surface expression of FasL by using flow cytometry. d Znrf1 -deficient macrophages were reconstituted with doxycycline-inducible wild-type ZNRF1, ZNRF1(C184A) mutant, or vector control. Following LPS stimulation (100 ng/mL, 24 h), co-immunoprecipitation was performed with anti-Munc18-2 and immunoblotting for Syntaxin-3 and Munc18-2. Syntaxin-3/Munc18-2 binding ratios are indicated. e Surface FasL expression was analyzed by flow cytometry in reconstituted macrophages from ( d ) under vehicle and LPS-treated conditions.

Article Snippet: FasL cDNA, obtained from Origene (Rockville, MD), was polymerase chain reaction–amplified and cloned into a C-terminally FLAG-tagged pLKO-AS2 lentiviral vector.

Techniques: Immunoprecipitation, Control, Western Blot, Confocal Microscopy, Staining, Fluorescence, Knockdown, Expressing, Flow Cytometry, Mutagenesis, Plasmid Preparation, Binding Assay

Journal: Cell Death & Disease

Article Title: ZNRF1 deficiency disrupts Fas ligand trafficking and immune balance

doi: 10.1038/s41419-026-08566-8

Figure Lengend Snippet: Schematic illustration of FasL trafficking and T cell apoptosis in wild-type (WT, left) versus Znrf1 -deficient (right) macrophages. In WT macrophages, ZNRF1 promotes the interaction between Munc18-2 and Syntaxin-3 (Stx3), enabling efficient delivery of FasL-containing vesicles to the plasma membrane and triggering FasL-mediated apoptosis of activated T lymphocytes. In Znrf1 -deficient macrophages, disruption of the Munc18-2/Syntaxin-3 interaction impairs FasL vesicle trafficking and surface expression, resulting in defective Fas-mediated T cell apoptosis and immune dysregulation. The lower panel depicts the in vivo allogeneic transfusion model, in which C57BL/6 Znrf1 f/f or Znrf1 myeΔ recipients received BALB/c whole blood via intravenous injection.

Article Snippet: FasL cDNA, obtained from Origene (Rockville, MD), was polymerase chain reaction–amplified and cloned into a C-terminally FLAG-tagged pLKO-AS2 lentiviral vector.

Techniques: Clinical Proteomics, Membrane, Disruption, Expressing, In Vivo, Injection

Journal: PLOS One

Article Title: Differential expression and regulation of FASLG by miR-5195/miR-3941 in age-related hearing loss

doi: 10.1371/journal.pone.0331661

Figure Lengend Snippet: (A). Heat map illustrating differential expression data from RNA-Seq analysis. Heatmap of FASLG gene expression, represented as fold changes in mRNA levels, was generated for the ARHL (n = 12) and NH (n = 12); (B). Expression of FASLG was analyzed using RT-qPCR and confirmed through WB (NH: n = 12, ARHL: n = 14). (C). Expression of miR-5195 was detected using RT-qPCR in WB (NH: n = 6, ARHL: n = 5). (D). Expression of miR-3941 was detected using RT-qPCR in WB (NH: n = 14, ARHL: n = 12). U6 snRNA was used as an internal control (** p < 0.01). Abbreviations used are as follows: Normal hearing group (NH), Age-related hearing loss (ARHL), Whole blood (WB), Fas ligand ( FASLG ).

Article Snippet: For the transfection experiments, the FASLG Human 3’ UTR Clone (#SC210991, OriGene, Rockville, Maryland, USA), miR-5195 (HmiR1468 – MR04, Genecopoeia, Rockville, Maryland, USA), and miR-3941 (HmiR1052 – MR04, Genecopoeia, Rockville, Maryland, USA) were used.

Techniques: Quantitative Proteomics, RNA Sequencing, Gene Expression, Generated, Expressing, Quantitative RT-PCR, Control

Journal: PLOS One

Article Title: Differential expression and regulation of FASLG by miR-5195/miR-3941 in age-related hearing loss

doi: 10.1371/journal.pone.0331661

Figure Lengend Snippet: (A). Identification of 3′-UTR sequences in the FASLG gene predicted to include binding sites for miR-3941; (B). Identification of 3′-UTR sequences in the FASLG gene predicted to harbor binding sites for miR-5195-5p; (C). Dual luciferase reporter assay results for miR-3941, miR-5195, and their corresponding target 3’-UTR of FASLG in HEI-OC-1 cells; (D). Dual luciferase assay results of miR-3941, miR-5195 and their corresponding target 3’-UTR of FASLG in HeLa cell; The significant differences between FASLG and the miRNA target group are represented as ** p < 0.01. Abbreviations used are as follows: Fas ligand ( FASLG ), House Ear Institute-Organ of Corti 1 (HEI-OC-1).

Article Snippet: For the transfection experiments, the FASLG Human 3’ UTR Clone (#SC210991, OriGene, Rockville, Maryland, USA), miR-5195 (HmiR1468 – MR04, Genecopoeia, Rockville, Maryland, USA), and miR-3941 (HmiR1052 – MR04, Genecopoeia, Rockville, Maryland, USA) were used.

Techniques: Binding Assay, Luciferase, Reporter Assay

Journal: PLOS One

Article Title: Differential expression and regulation of FASLG by miR-5195/miR-3941 in age-related hearing loss

doi: 10.1371/journal.pone.0331661

Figure Lengend Snippet: (A). The light blue rectangular node represents the FASLG gene, while the light blue circular nodes represent the 35 selected miRNAs. The nodes with red text indicate the two miRNAs of interest, miR-3941 and miR-5195-5p. The arrows connecting FASLG to the miRNAs indicate that the thickness of the arrow corresponds to the context++ score of the miRNA, with thicker arrows representing lower scores. (B). We utilized blood samples from individuals with ARHL and found that A notable rise in FASLG expression was observed. Additionally, we identified miR-3941 and miR-5195 as potential regulators of FASLG expression. These miRNAs may bind to the 3’ UTR of the elevated FASLG , leading to down-regulation of gene expression. Abbreviations used are as follows: Fas ligand ( FASLG ), Age-related hearing loss (ARHL), Fas receptor (FAS-R), 3’untranslated region (3’UTR).

Article Snippet: For the transfection experiments, the FASLG Human 3’ UTR Clone (#SC210991, OriGene, Rockville, Maryland, USA), miR-5195 (HmiR1468 – MR04, Genecopoeia, Rockville, Maryland, USA), and miR-3941 (HmiR1052 – MR04, Genecopoeia, Rockville, Maryland, USA) were used.

Techniques: Expressing, Gene Expression

Journal: Biological & pharmaceutical bulletin

Article Title: Lx2-32c, a novel taxane derivative, exerts anti-resistance activity by initiating intrinsic apoptosis pathway in vitro and inhibits the growth of resistant tumor in vivo.

doi: 10.1248/bpb.b12-00513

Figure Lengend Snippet: Fig. 5. Effect of Lx2-32c on the Expression of Fas/FasL and Caspase-8 in MX-1/T Cells

Article Snippet: The membranes were blocked with 5% fat-free dry milk in TBST for 2 h, and probed overnight at 4°C with rabbit monoclonal antibodies against caspase-3, caspase-8, caspase-9, poly(ADP-ribose) polymerase (PARP), Fas, FasL, Bax and Bcl-2 (CST, U.S.A.), respectively.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Candidate protein biomarkers in pancreatic neuroendocrine neoplasms grade 3

doi: 10.1038/s41598-020-67670-7

Figure Lengend Snippet: PanNEN G3 immunostained for FASLG. ( a ), ( b ) and ( c ) FASLG immunoreactivity in membrane of tumour cells. ( d ) FASLG immunoreactive immune cells infiltrating a non-immunoreactive tumour. Scale bar 100 µm.

Article Snippet: The primary rabbit polyclonal antibody against FAS ligand (FASLG) (HPA054959, Atlas Antibodies, Stockholm, Sweden) was diluted in 1:800 UltraAb Diluent (Thermo Fisher Scientific) followed by incubation for 30 min at room temperature (RT).

Techniques: Membrane

Journal: Scientific Reports

Article Title: Candidate protein biomarkers in pancreatic neuroendocrine neoplasms grade 3

doi: 10.1038/s41598-020-67670-7

Figure Lengend Snippet: FASLG expression in well- and poorly differentiated tumour samples.

Article Snippet: The primary rabbit polyclonal antibody against FAS ligand (FASLG) (HPA054959, Atlas Antibodies, Stockholm, Sweden) was diluted in 1:800 UltraAb Diluent (Thermo Fisher Scientific) followed by incubation for 30 min at room temperature (RT).

Techniques: Expressing