mca2142 rrid ab 324562  (Bio-Rad)

Journal: iScience

Article Title: Plasma extracellular vesicle modulate immune cell transcriptional responses following acute myocardial infarction

doi: 10.1016/j.isci.2026.114665

Figure Lengend Snippet: Characterization of plasma EV in MI patients EVs were isolated from the plasma of MI patients at hospital presentation and at 3-month follow-up. (A) EV-Array analysis of canonical EV-associated markers shows significant enrichment at MI presentation ( n = 17 patients paired at both timepoints). (B) EV-cell-associated markers were also enriched at presentation with MI. White in the heatmaps denotes the baseline (normalized to 1), corresponding to the 3-month follow-up value. (C) Total EV concentrations measured by NTA of SEC and ultracentrifugation-purified plasma EVs. (D and E) Spaghetti plots showing EV size and particle concentration profiles at the individual patient level across the two timepoints ( n = 19 patients paired at both timepoints). (E) Western blotting confirmed the presence of EV markers (CD9 and Syntenin-1), absence of cellular contaminants (GM130, ATP5A, and Histone H3), and depletion of the plasma Apo B. (F) TEM images confirmed typical cup-shaped EV morphology. Scale bars, 500 nm or 100 nm. (G and H) Single-particle interferometric reflectance imaging (NanoView) confirmed tetraspanin-positive EVs (CD9, CD63, and CD81), (H) with associated size and concentration profiles ( n = 9–10 at both timepoints). EV marker values at MI presentation were normalized to each patient’s own 3-month follow-up value. In (G), data are presented as group means ± SD. Statistical analysis: (A–C) Wilcoxon matched-pairs signed-rank test; (G) two-way ANOVA with Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: CD63 , Biorad , MCA2142; RRID: AB_324562.

Techniques: Clinical Proteomics, Isolation, Purification, Concentration Assay, Western Blot, Single Particle, Imaging, Marker

Journal: iScience

Article Title: Plasma extracellular vesicle modulate immune cell transcriptional responses following acute myocardial infarction

doi: 10.1016/j.isci.2026.114665

Figure Lengend Snippet: Enrichment of miRNA-320b in endothelial cell-derived EV following inflammatory stimulation HUVECs were cultured under control (black) or TNF-α-stimulated (red) conditions to model endothelial inflammation. (A) Total and viable cell counts at study endpoint confirm similar cell numbers across conditions. (B) Soluble VCAM-1 levels in supernatants were significantly increased following TNF-α stimulation, confirming proinflammatory activation. (C and D) Single-particle interferometric reflectance imaging (NanoView) detected CD9, CD63, and CD81-positive EVs in both conditions. Scale bars, 10 μm. (D) Total captured EV concentrations were significantly higher in TNF-α-stimulated cultures. (E) Average particle diameters of captured EVs did not differ significantly. (F) NTA showed elevated particle concentrations in EV preparations from TNF-α-treated HUVECs. (G) Size distribution profiles of EVs from control and TNF-α conditions. (H) EV counts were normalized to total and live cell numbers, confirming increased release under inflammatory stimulation. (I) Western blot analysis of HUVEC-derived EVs confirmed enrichment of canonical EV markers (TSG101, Syntenin-1, and CD9), absence of cellular contaminant GM130, and presence of VCAM-1 specifically in TNF-α-derived EVs. (J) TEM images showed characteristic EV morphology; scale bars, 1 μm or 500 nm. (K) RT-qPCR showed no change in HUVEC cellular miRNA-320b levels between conditions. (L) Significant enrichment of miRNA-320b was detected in EVs from TNF-α-stimulated HUVECs. miRNA expression was normalized to UniSp6 spike-in control, because endogenous controls (e.g., miR-103a-3p) were undetectable in EV samples. Data are presented as group means ± SD. N = 4 biological replicates per group. Statistical analysis: (B, F, L) unpaired Student’s t test; (D, H) two-way ANOVA with Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: CD63 , Biorad , MCA2142; RRID: AB_324562.

Techniques: Derivative Assay, Cell Culture, Control, Activation Assay, Single Particle, Imaging, Western Blot, Quantitative RT-PCR, Expressing