fasl cdna  (OriGene)

Journal: Journal of Neurology

Article Title: Soluble TIM-3 and galectin-9 serve as additional diagnostic biomarkers in primary central nervous system lymphoma

doi: 10.1007/s00415-026-13791-4

Figure Lengend Snippet: Accuracy of CSF diagnostic markers for PCNSL. Boxplots depicted the distribution of CSF IL-10 ( A ), CXCL13 ( B ), IL-2RA ( C ), FasL ( D ), TIM-3 ( E ), and galectin-9 ( F ) levels in PCNSL, neuroimmunological diseases, other brain tumors, and iNPH patients. Median values were represented by the horizontal bar, IQR by boxes, 1.5 × IQR by whiskers, and individual values by dots. IL-10 and IL-2RA are shown on a logarithmic scale due to the highly skewed distribution with a large proportion of zero or low values and a small number of extremely high values. The y -axis is displayed on a logarithmic scale after log10 transformation of IL-10 and IL-2RA concentrations with the addition of a constant of 1, while tick labels indicate the original values in pg/mL. **, adjusted p < 0.01. ***, adjusted p < 0.001. n.s., not significant. Holm-adjusted Dunn’s tests were used to compare PCNSL with the other groups. G–I ROC curves showed the diagnostic performance of CSF IL-10, CXCL13, IL-2RA, FasL, TIM-3, and galectin-9 for distinguishing PCNSL from different disease groups. The corresponding AUC values were shown. G PCNSL versus all other diseases. H PCNSL versus neuroimmunological diseases. (I) PCNSL versus other brain tumors. AUC values were shown in the legends of each panel AUC, area under the curve; CSF, cerebrospinal fluid; CXCL13, C–X–C motif chemokine ligand 13; FasL, Fas ligand; IL-2RA, interleukin-2 receptor chain alpha; IL-10, interleukin-10; iNPH, idiopathic normal pressure hydrocephalus; IQR, interquartile range; PCNSL, primary central nervous system lymphoma; ROC, Receiver operating characteristic; TIM-3, T-cell immunoglobulin and mucin domain 3

Article Snippet: FasL + Galectin-9 , 68.5 , 76.8 , 0.961 , 0.269 , 0.631 , .

Techniques: Diagnostic Assay, Transformation Assay

Journal: Journal of Neurology

Article Title: Soluble TIM-3 and galectin-9 serve as additional diagnostic biomarkers in primary central nervous system lymphoma

doi: 10.1007/s00415-026-13791-4

Figure Lengend Snippet: Correlations and clustering of diagnostic markers in PCNSL. Scatter plots and correlation analyses showed correlations between IL-10 and CXCL13 ( A ), IL-10 and IL-2RA ( B ), IL-10 and FasL ( C ), IL-10 and TIM-3 ( D ), IL-10 and galectin-9 ( E ), CXCL13 and IL-2RA ( F ), CXCL13 and FasL ( G ), CXCL13 and TIM-3 ( H ), CXCL13 and galectin-9 ( I ), IL-2RA and FasL ( J ), IL-2RA and TIM-3 ( K ), IL-2RA and galectin-9 (L), FasL and TIM-3 ( M ), FasL and galectin-9 ( N ), and TIM-3 and galectin-9 ( O ). Spearman’s rank correlation was used to measure the degree of correlation between CSF diagnostic markers. P Correlation matrix showed Spearman’s rank correlation coefficients between CSF diagnostic markers in PCNSL patients. Q Hierarchical clustering of CSF diagnostic markers in PCNSL patients based on Spearman’s rank correlation coefficients. Clustering was performed using 1 − Spearman’s correlation coefficient as the distance metric with the complete linkage method. CSF, cerebrospinal fluid; CXCL13, C–X–C motif chemokine ligand 13; FasL, Fas ligand; IL-2RA, interleukin-2 receptor chain alpha; IL-10, interleukin-10; PCNSL, primary central nervous system lymphoma; TIM-3, T-cell immunoglobulin and mucin domain 3

Article Snippet: FasL + Galectin-9 , 68.5 , 76.8 , 0.961 , 0.269 , 0.631 , .

Techniques: Diagnostic Assay

Journal: Journal of Neurology

Article Title: Soluble TIM-3 and galectin-9 serve as additional diagnostic biomarkers in primary central nervous system lymphoma

doi: 10.1007/s00415-026-13791-4

Figure Lengend Snippet: Comparison of PFS in PCNSL according to diagnostic marker levels. Kaplan–Meier curves for PFS in PCNSL patients were compared according to CSF IL-10 ( A ), CXCL13 ( B ), IL-2RA ( C ), FasL ( D ), TIM-3 ( E ), and galectin-9 ( F ) levels. PFS was compared using the log-rank test. CSF, cerebrospinal fluid; CXCL13, C–X–C motif chemokine ligand 13; FasL, Fas ligand; IL-2RA, interleukin-2 receptor chain alpha; IL-10, interleukin-10; PCNSL, primary central nervous system lymphoma; PFS, progression-free survival; TIM-3, T-cell immunoglobulin and mucin domain 3

Article Snippet: FasL + Galectin-9 , 68.5 , 76.8 , 0.961 , 0.269 , 0.631 , .

Techniques: Comparison, Diagnostic Assay, Marker

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