Journal: Life science alliance

Article Title: Megakaryocytes possess a STING pathway that is transferred to platelets to potentiate activation.

doi: 10.26508/lsa.202302211

Figure Lengend Snippet: Figure 1. STING stimulation of megakaryocytes induces a type-I interferon response. (A) Brightfield images of megakaryocytes isolated using anti-CD41 magnetic beads at day 4 of in vitro differentiation. Enlargement shows the production of proplatelets (black arrows) by mature megakaryocytes. Scale bar: 30 µm. (B, C) Cytokine production analysis by ELISA of megakaryocytes stimulated with DMXAA for 5 h (B) and 24 h (C) showing a significant increase in IFNB and CCL5 cytokines, n = 4 independent experiments; each dot represents data from one independent experiment. The graph shows data ± STD, and data were analysed by unpaired two-tailed t tests, *P < 0.05. (D, E) Representative immunofluorescence images of megakaryocytes taken by confocal microscopy showing the localisation of cGAS or STING (red), ERp57 or lamin A/C (green), and chromatin (blue), representative of n = 3 independent experiments. Scale bars: 20 µm (D) and 15 µm (E). (F) Representative immunofluorescence images of CD41-positive (magenta for CD41+ and blue for DNA) and CD41-negative (only blue) cells taken by confocal microscopy. Representative images of more than n = 3 independent experiments. Scale bars: 30 µm.

Article Snippet: Cells were then labelled with 10 μl anti-CD41 antibody (coupled to Biotin or APC—Miltenyi Biotec) for 10 min at 4°C, then washed with 2 ml of isolation buffer, and centrifuged at 300g for 10 min at 4°C, twice before the addition of 20 μl of Anti-Biotin/APC MicroBeads (Miltenyi Biotec) and 70 μl of isolation buffer for 15 min at 4°C.

Techniques: Isolation, Magnetic Beads, In Vitro, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Confocal Microscopy

Journal: International journal of molecular sciences

Article Title: Pulmonary Biodistribution of Platelet-Derived Regenerative Exosomes in a Porcine Model.

doi: 10.3390/ijms25052642

Figure Lengend Snippet: Figure 1. Exosome characterization of PEP and membrane labeling by lipophilic dye. (A) NanoSight analysis of PEP demonstrating distribution of particle size ± standard deviation (gray shading). (n = 3). (B) Transmission electron microscopy image of PEP with scale bar 2 µm. (C) Western blot demonstrating presence of hallmark exosomal proteins CD9, CD63, and Flotillin, as well as platelet integrin CD41. (D) Immunocytochemistry of human umbilical vein endothelial cells following treatment with DiI-labeled PEP or DiI alone (control) in red, counterstained with phalloidin (green) and DAPI (blue), scale bar 20 µm. (E) Quantification (n = 5) of mean integrated density DiI per nuclei (±SD) for Figure 1D. ** p < 0.001 using Mann-Whitney two-tailed t-test. (F) Xenogen image demonstrating fluorescent signal detected in unlabeled PEP, DiI, and PEP + DiI. (G) Xenogen image demonstrating fluorescent signal detected in unlabeled PEP, DiR, and PEP + DiR.

Article Snippet: Primary antibodies included CD63 (1:1000, R&D Systems, Minneapolis, MN, USA, #MAB50482), Flotillin (1:1000, Abcam, Waltham, MA, USA, #ab133497), CD9 (1:1000, Cell Signaling Technology, Danvers, MA, USA, #13403S), CD41 (1:1000, NovusBiologicals, Centennial, CO, USA, #NBP1-84581), GAPDH (1:2000, Cell Signaling Technology, 2118S), and actin (1:1000, LI-COR, #926-42212).

Techniques: Membrane, Labeling, Standard Deviation, Transmission Assay, Electron Microscopy, Western Blot, Immunocytochemistry, Control, MANN-WHITNEY, Two Tailed Test

Journal: iScience

Article Title: Inflammation-responsive biomimetic nanoparticles with epigallocatechin-3-gallate for acute lung injury therapy via autophagy enhancement

doi: 10.1016/j.isci.2025.112318

Figure Lengend Snippet:

Article Snippet: CD41 Antibody , Proteintech , Cat # 24552-1-AP; RRID: AB_2879604.

Techniques: Recombinant, Software

Journal: Physiological Research

Article Title: The Lung Is Not a Primary Site of Platelet Biogenesis

doi: 10.33549/physiolres.935477

Figure Lengend Snippet: Megakaryocytes in human lungs were detected by single-cell RNA sequencing. Platelet factor 4 (PF4) expression (blue) based on 10X Chromium- ( A ) and SmartSeq2 (SS2)-based ( B ) single-cell RNA sequencing. ( C ) The percentages of PF4 + , ITGA2B + , ITGB3 + , and mean positive cells based on the indicated methods.

Article Snippet: Sections were deparaffinized, incubated with 3 % H 2 O 2 for 30 min to block endogenous peroxidase activity, retrieved with Tris-EDTA solution (pH 9.0) in a pressure cooker, blocked with 10 % serum, and then incubated with anti-PF4 (A3694), ITGA2B (A11490), and ITGB3 (A19073) antibodies (ABclonal, Wuhan, China) followed by horseradish peroxidase conjugate secondary antibodies (ZSGB-BIO, Beijing).

Techniques: RNA Sequencing, Expressing