mouse pf4 elisa kit  (Elabscience Biotechnology)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: TLR8 agonists remodel the tumor immune microenvironment through PF4-dependent T cell recruitment and ancillary mechanisms

doi: 10.1007/s00262-026-04329-8

Figure Lengend Snippet: TLR8 agonist activates PF4-CXCR3 pathway to shape the TIME. A t-SNE plot showing the distribution of all cells from the control and agonist-treated groups combined ( n = 15,745 cells). B t-SNE plot depicting the distribution of distinct immune cell types. C Dot plot displaying the marker genes used to define each immune cell population. D Density plot showing the distribution of Tlr8-expressing cells across the t-SNE map. E Violin plot illustrating Tlr8 expression levels across different immune cell types. F Circle plot depicting the strength of Pf4–Cxcr3 receptor–ligand interactions between distinct cell types in the control and motolimod-treated groups. G Differential gene expression profiles of neutrophils, macrophages, cDC2, and CD8 + T cells. H Bubble plot comparing changes in specific receptor–ligand interactions between the control and motolimod-treated groups, focusing on macrophage-to-cCD4 + T signaling, cDC2-to-cCD4 + T signaling, and CD8 + T-to-cCD4 + T signaling. I Violin plot showing differential expression of Cxcr3 in cCD4 + T and Tregs, analyzed using the Wilcoxon rank-sum test. ns, not significant; ** P < 0.01

Article Snippet: Pf4 levels in culture supernatants were quantified using Mouse PF4 ELISA Kit (Elabscience, #E-EL-M3080) according to the manufacturer’s instructions.

Techniques: Control, Marker, Expressing, Gene Expression, Quantitative Proteomics

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: TLR8 agonists remodel the tumor immune microenvironment through PF4-dependent T cell recruitment and ancillary mechanisms

doi: 10.1007/s00262-026-04329-8

Figure Lengend Snippet: TLR8 agonist induces PF4 expression through NF-κB signaling pathway. A , B qRT-PCR analysis of Pf4 expression in mouse and human macrophages in the motolimod-treated and control groups ( n = 3). C , D Luminescence assay showing NF-κB pathway activation in mouse and human macrophages following motolimod stimulation ( n = 3). E , F qRT-PCR analysis of Rela expression in Rela -knockdown and control mouse and human macrophages with or without motolimod treatment ( n = 3). G , H Western blotting showing p65 protein levels in RELA -knockdown and control mouse and human macrophages with or without motolimod treatment. I , J qRT-PCR analysis of Pf4 / PF4 expression in Rela / RELA -knockdown and control mouse and human macrophages with or without motolimod treatment ( n = 3). K ELISA quantification of Pf4 levels in the culture supernatant from Rela -knockdown and control mouse macrophages with or without motolimod treatment ( n = 3). Data represent the mean ± SEM. Statistical comparisons were performed using Student’s t-tests. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Pf4 levels in culture supernatants were quantified using Mouse PF4 ELISA Kit (Elabscience, #E-EL-M3080) according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Control, Luminescence Assay, Activation Assay, Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: TLR8 agonists remodel the tumor immune microenvironment through PF4-dependent T cell recruitment and ancillary mechanisms

doi: 10.1007/s00262-026-04329-8

Figure Lengend Snippet: PF4 remodels the TIME and suppresses tumor growth. A , B Tumor growth curves and terminal tumor weights in C57BL/6 mice bearing AKR or MC38 tumors with or without Pf4 overexpression, treated with or without motolimod administered every three days ( n = 6). C Percentage of CD45 + cells among live cells in AKR and MC38 tumor models ( n = 6). D Percentage of cCD4 + T cells within the CD45 + population in AKR and MC38 tumor models ( n = 6). E Percentage of CD8 + T cells within the CD45 + population in AKR and MC38 tumor models ( n = 6). F Percentage of Tregs within the CD45 + population in AKR and MC38 tumor models ( n = 6). G Ratios of cCD4 + T cells to Tregs in AKR and MC38 tumor models ( n = 6). H Ratios of CD8 + T cells to Tregs in AKR and MC38 tumor models ( n = 6). I Representative immunofluorescence staining of CD8α (red) and DAPI (blue) in MC38-Vector or MC38-Pf4-OE tumor sections with or without motolimod treatment. Scale bar: 50 μm. J Quantification of CD8α + positive cell ratio (%) relative to total DAPI + nucleated cells per section ( n = 6). Data represent the mean ± SEM. Statistical comparisons were performed using Student’s t-tests. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Pf4 levels in culture supernatants were quantified using Mouse PF4 ELISA Kit (Elabscience, #E-EL-M3080) according to the manufacturer’s instructions.

Techniques: Over Expression, Immunofluorescence, Staining, Plasmid Preparation

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Induced pluripotent stem cell-derived platelets kill multidrug-resistant Staphylococcus aureus via Toll-like receptor 2-MyD88 signaling and immunoglobulin G/FcγRIIA engagement

doi: 10.1016/j.rpth.2026.103374

Figure Lengend Snippet: Induced pluripotent stem cell (iPSC)-derived platelets (iPSC-PLTs) exhibit antimultidrug-resistant Staphylococcus aureus (MRSA) activity. (A) Schema of the MRSA-killing assay. Colony formation units on Todd-Hewitt broth (THB) agar plates were counted. (B) MRSA-killing assay using 3 iPSC-PLT clones (Sev2, NIH5, and M35-1) and healthy peripheral blood (PB) platelets (PLTs). MRSA killing was evaluated by quantifying MRSA colony numbers relative to the MRSA-only control as a percentage. (C–E) SeV2 iPSC-PLTs with or without 2-hour coincubation with MRSA were subject to flow cytometry analysis for the (C) expression of CD62P, CD63, and CD107a and the binding of PAC-1 and annexin V, (D) enzyme-linked immunosorbent assay for the release of platelet factor 4 (PF4) and β-thromboglobulin (β-TG), and (E) chemiluminescent enzyme immunoassay for the release of sCLEC-2. (F) MRSA-killing assay using SeV2 iPSC-PLTs pretreated with gradient concentrations of tirofiban, aspirin, BPTU, or ticagrelor. Compiled data of 3 independent experiments. Data are presented as means ± SEM. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001, ADP, adenosine diphosphate; ns, not significant. MFI, mean fluorescence intensity; sCLEC-2, soluble C-type lectin-like receptor 2; TRAP-6, thrombin receptor activator peptide 6.

Article Snippet: Platelet factor 4 (PF4) and β-thromboglobulin (β-TG) were quantified using commercial ELISA kits (R&D Systems, #DY795 and PeproTech, #900-K40) according to the manufacturer’s instructions, with samples diluted 1:50 in phosphate-buffered saline.

Techniques: Derivative Assay, Activity Assay, Clone Assay, Control, Flow Cytometry, Expressing, Binding Assay, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: bioRxiv

Article Title: Platelet C5aR1 Aggravates Myocardial Infarction through Platelet–Neutrophil Interactions and CXCL4-Dependent NET Release

doi: 10.64898/2026.01.12.699090

Figure Lengend Snippet: (A) Representative single-plane confocal immunofluorescence images of fixed and permeabilized resting wild-type (WT) and C5aR1-deficient platelets stained for P-selectin (CD62P, green) and CXCL4 (red), illustrating altered α-granule organization. (B) Quantification of intracellular α-granule content per platelet, expressed as total area of P-selectin–positive granules and colocalized P-selectin/CXCL4 granules, measured by confocal microscopy. Data are shown as individual platelet values pooled from n = 3 independent experiments. Outliers were identified and removed using the ROUT method (Q = 1%) prior to analysis. (C) Flow cytometric analysis of platelet surface P-selectin expression 24 h after myocardial infarction following ex vivo stimulation of whole blood with 100 nM phorbol 12-myristate 13-acetate (PMA), expressed as geometric mean fluorescence intensity (GMFI) of CD42b⁺ platelets. (D) Flow cytometric analysis of platelet integrin GPIIb/IIIa activation under the same conditions, expressed as percentage of activated GPIIb/IIIa among CD42b⁺ platelets. (E) Plasma CXCL4 concentrations after MI in Pf4^cre+^ C5aR1^fl/fl^ mice and Cre-negative littermate controls. (F) Schematic of the in vitro platelet–neutrophil co-incubation assay. WT or C5aR1-deficient platelets were stimulated with C5a and co-incubated with neutrophils, followed by confocal immunofluorescence staining for myeloperoxidase (MPO), citrullinated histone H3 (H3Cit), and DNA (DAPI) to assess NET formation, in the presence or absence of low-dose heparin or recombinant CXCL4 (rCXCL4). (G) Representative immunofluorescence images of neutrophils after co-incubation, stained for MPO (red), H3Cit (green), and DNA (DAPI, blue), illustrating NET formation under the indicated conditions (see Supplementary Figure 13 for neutrophil-intrinsic and platelet-mediated control conditions). (H) Quantification of NET formation expressed as percentage of H3Cit⁺ neutrophils. Data are shown as mean ± SD unless otherwise indicated; each dot represents one biological replicate or mouse, as indicated. For α-granule analyses (A–B), data are shown as individual platelet values pooled from n = 3 independent experiments and analyzed using two-tailed unpaired t-tests following ROUT-based outlier exclusion (Q = 1%). Flow cytometry data (C–D) were analyzed using one-way ANOVA across time points and genotypes (see also Supplementary Figures for day 14 analyses). Plasma CXCL4 measurements (E) were analyzed using two-tailed unpaired t-tests. NET formation assays (H) were analyzed using one-way ANOVA with appropriate post hoc correction. P < 0.05, P < 0.01, P < 0.001, P < 0.0001. Scale bars, 2 µm (A) and 20 µm (G).

Article Snippet: CXCL4 concentrations were measured in plasma samples using a mouse CXCL4 (platelet factor 4) Quantikine ELISA kit (R&D Systems, MCX400) according to the manufacturer’s instructions.

Techniques: Immunofluorescence, Staining, Confocal Microscopy, Expressing, Ex Vivo, Fluorescence, Activation Assay, Clinical Proteomics, In Vitro, Incubation, Recombinant, Control, Two Tailed Test, Flow Cytometry