Journal: Journal of Extracellular Biology

Article Title: Monocyte derived large extracellular vesicles in polytrauma

doi: 10.1002/jex2.70005

Figure Lengend Snippet: List of antibodies and isotypes used for cell‐ and cell derived large EV flow cytometric analysis. The antibodies were titrated against their matching isotype controls prior use and applied according to the manufacturers’ instructions.

Article Snippet: CD14 (mouse IgG2a) , VioBlue , 130‐094‐364 , Miltenyi Biotec , 0.01 , 1:50.

Techniques: Derivative Assay

Journal: Journal of Extracellular Biology

Article Title: Monocyte derived large extracellular vesicles in polytrauma

doi: 10.1002/jex2.70005

Figure Lengend Snippet: Antibody validation on cultured THP‐1 cells for EV detection and general used large EV gating strategy . (a) In vitro human THP‐1 monocyte cells were treated with PMA alone or PMA & LPS (each n ≥ 3), increasing CD14 expression. (b) Expression of CD14 on surface of AnnV + large EVs (lEVs) derived from non‐stimulated THP‐1 cells and from PMA alone or PMA & LPS stimulated THP‐1 cells. AnnV is used as a general large EV marker. Values given as mean with SEM. Three column statistical analysis was done by Kruskal–Wallis test including Dunn's post hoc test for multiple comparisons (*, p < 0.05; **, p < 0.01; ***, p < 0.001). (c) Thresholds testing for cell culture and serum derived large EV detection and gating. Prior use of flow cytometry gates sensitivity of used flow cytometer was set taking advantage of 1000, 500 and not detectable 200 nm beads. (d) Gating strategy for the detection of CD14‐APC on AnnV + large EVs controlled by matching REA isotype control.

Article Snippet: CD14 (mouse IgG2a) , VioBlue , 130‐094‐364 , Miltenyi Biotec , 0.01 , 1:50.

Techniques: Biomarker Discovery, Cell Culture, In Vitro, Expressing, Derivative Assay, Marker, Flow Cytometry, Control

Journal: Journal of Extracellular Biology

Article Title: Monocyte derived large extracellular vesicles in polytrauma

doi: 10.1002/jex2.70005

Figure Lengend Snippet: Semi quantification of human serum derived AnnV + CD14 + large EVs . (a) Workflow SOP for isolation of large EVs (lEVs) from 1 mL of human serum collected from polytrauma patients (ISS > 15, at day 0, at day 1 and day 7 post trauma). (b) AnnV + CD14 + large EVs counts in small EVs in polytrauma (ISS > 15) with internal organ damage (polytrauma OD) versus polytrauma (ISS ≥ 15) without internal organ damage (polytrauma w/o OD) upon admission (day 0). (c) AnnV + CD14 + large EVs counts in polytrauma (ISS > 15) polytrauma OD versus polytrauma w/o OD (ISS > 15) 24 h post trauma (day 1). (d) Day 1, corresponding AUROC, sensitivity, specificity, and cut‐off value, respectively. (e) AnnV + CD14 + large EVs counts in small EVs in polytrauma (ISS ≥ 15) polytrauma OD versus polytrauma w/o OD (ISS ≥ 15) 7 days post trauma (day 7). Values given as median with 95% CI. Three column statistical analysis was done by Kruskal–Wallis test including Dunn's post hoc test for multiple comparisons (*, p < 0.05; **, p < 0.01; ***, p < 0.001; not significant, n as indicated). Horizontal dotted line in panels B, C and E indicates calculated associated cut‐off (summarized in Table ).

Article Snippet: CD14 (mouse IgG2a) , VioBlue , 130‐094‐364 , Miltenyi Biotec , 0.01 , 1:50.

Techniques: Derivative Assay, Isolation

Journal: Journal of Extracellular Biology

Article Title: Monocyte derived large extracellular vesicles in polytrauma

doi: 10.1002/jex2.70005

Figure Lengend Snippet: AnnV + CD14 + large EVs kinetics at day 1 and day 7 in polytrauma (ISS > 15) and correlation with the Injury Severity Score (ISS) . (a) depicted are pairs of AnnV + CD14 + large EVs (lEVs) measured at day 1 and day 7 of enrolled polytrauma (ISS > 15). Values given as median. Statistical significance was assessed by two tailed paired t test (*, p < 0.05; **, p < 0.01; ***, p < 0.001). (b) Spearman ( r sp ) correlation between percentages of AnnV + CD14 + large EVs and ISS ( n = 24). p and r sp values as indicated.

Article Snippet: CD14 (mouse IgG2a) , VioBlue , 130‐094‐364 , Miltenyi Biotec , 0.01 , 1:50.

Techniques: Two Tailed Test

Journal: Journal of Extracellular Biology

Article Title: Monocyte derived large extracellular vesicles in polytrauma

doi: 10.1002/jex2.70005

Figure Lengend Snippet: Summarized data FACS of AnnV + CD14 + large EVS in polytrauma including associated SEM, percentage change and statistics at day 1.

Article Snippet: CD14 (mouse IgG2a) , VioBlue , 130‐094‐364 , Miltenyi Biotec , 0.01 , 1:50.

Techniques:

Journal: Journal of Extracellular Biology

Article Title: Monocyte derived large extracellular vesicles in polytrauma

doi: 10.1002/jex2.70005

Figure Lengend Snippet: List of antibodies used for human derived large EV surface marker analysis. The antibody was added according to manufacturer's recommendations and titrated against the matching isotype control prior use.

Article Snippet: CD14 (mouse IgG2a) , VioBlue , 130‐094‐364 , Miltenyi Biotec , 0.01 , 1:50.

Techniques: Derivative Assay, Marker, Control

Journal: iScience

Article Title: Clonal hematopoiesis is associated with cardiovascular events in patients with stable coronary artery disease

doi: 10.1016/j.isci.2024.109472

Figure Lengend Snippet:

Article Snippet: CD14-phycoerythrin, clone REA599 , Miltenyi Biotec, Bergisch Gladbach, Germany , Cat#130-110-519; RRID: AB_2655052.

Techniques: Enzyme-linked Immunosorbent Assay, Software, Flow Cytometry