Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Comparative phenotyping of surface markers and glycans in murine and human platelet-derived extracellular vesicles

doi: 10.1016/j.rpth.2026.103414

Figure Lengend Snippet: Surface glycans on l-PEVs in human and mice. The binding of fluorescein-conjugated RCA-1 (A), sWGA (B), SNA (C), MAL (D), ConA (E), and UEA lectins (F) to mouse and human l-PEVs was measured using flow cytometry. Data are represented as mean fluorescence intensity (MFI) in arbitrary units (a.u.). Orange, blue, and green lines indicate comparison between human and murine samples in resting, CRP + PAR1/4-AP stimulated (CT), or A23187 stimulated conditions, respectively. Quantification of carbohydrates in lysates of l-PEVs and platelets was performed using a microarray containing 95 lectins. (G) Comparison between total lectin binding to human and murine l-PEVs. (H) Comparison between total lectin binding to human and mouse platelets. Murine l-PEVs contained less carbohydrates, including galactose (I), N -acetylglucosamine (J), sialic acid (K), T-antigen core sugars (L), mannose (M), fucose (N), and N -acetylgalactosamine (O). Data are represented in relative fluorescence units after normalization of signal on positive and negative controls for protein binding to lectins. Data were obtained after IQR outlier test was performed. Data are mean ± SD; n = 8 to 12; ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001. ConA, concanavalin A; GlcNAc, β-galactose and N -acetylglucosamine; MAL, Maackia amurensis ; PEV, platelet-derived extracellular vesicle; RCA, Ricinus communis agglutinin; SNA, Sambucus nigra ; sWGA, succinylated wheat germ agglutinin; UEA, Ulex europaeus agglutinin.

Article Snippet: Ricinus communis agglutinin 1, succinylated wheat germ agglutinin, Sambucus nigra , Maackia amurensis , concanavalin A, and Ulex europaeus agglutinin (all fluorescein-conjugated) were from Vector Laboratories.

Techniques: Binding Assay, Flow Cytometry, Fluorescence, Comparison, Microarray, Protein Binding, Derivative Assay

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Comparative phenotyping of surface markers and glycans in murine and human platelet-derived extracellular vesicles

doi: 10.1016/j.rpth.2026.103414

Figure Lengend Snippet: Comparison of l-PEVs generated from BCs- and fresh blood-derived human platelets. (A) Platelets were isolated from either BCs or freshly drawn citrated human whole blood. l-PEVs from unstimulated and activated (CT, A23187) conditions were analyzed using flow cytometry. Concentration of generated l-PEVs (B) and their procoagulant properties (C, D) were assessed in Trucount tubes. (E) Percentage of CD42b + l-PEVs. (F) Percentage of CD62P + l-PEVs. (G) Percentage of l-GPVI + PEVs. Surface binding of RCA-1 (H), sWGA (I), SNA (J), MAL (K), ConA (L), and UEA lectins (M). Green line depicts comparison between A23187-stimulated samples. Data are mean ± SD; n = 6; ∗∗ P < .01. BC, buffy coat; ConA, concanavalin A; CRP, collagen-related peptide; GlcNAc, β-galactose and N -acetylglucosamine; MAL, Maackia amurensis ; MFI, mean fluorescence intensity; ns, not significant; PEV, platelet-derived extracellular vesicle; RCA, Ricinus communis agglutinin; SNA, Sambucus nigra ; sWGA, succinylated wheat germ agglutinin; UEA, Ulex europaeus agglutinin.

Article Snippet: Ricinus communis agglutinin 1, succinylated wheat germ agglutinin, Sambucus nigra , Maackia amurensis , concanavalin A, and Ulex europaeus agglutinin (all fluorescein-conjugated) were from Vector Laboratories.

Techniques: Comparison, Generated, Derivative Assay, Isolation, Flow Cytometry, Concentration Assay, Binding Assay, Fluorescence

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Comparative phenotyping of surface markers and glycans in murine and human platelet-derived extracellular vesicles

doi: 10.1016/j.rpth.2026.103414

Figure Lengend Snippet: Comparison of l-PEVs generated from platelets isolated from citrated or heparinized murine blood. (A) Platelets were isolated from citrated or heparinized murine whole blood. l-PEVs from unstimulated and activated (CT, A23187) conditions were analyzed using flow cytometry. Concentration of generated l-PEVs (B) and their procoagulant properties (C, D) were assessed in Trucount tubes. (E) Percentage of CD42b + l-PEVs. (F) Percentage of CD62P + l-PEVs. (G) Percentage of GPVI + l-PEVs. Surface binding of RCA-1 (H), sWGA (I), SNA (J), MAL (K), ConA (L), and UEA lectins (M). Orange line depicts the comparison between resting samples. Data are mean ± SD; n = 6; ∗ P < .05. ConA, concanavalin A; CRP, collagen-related peptide; GlcNAc, β-galactose and N -acetylglucosamine; MAL, Maackia amurensis ; MFI, mean fluorescence intensity; ns, not significant; PEV, platelet-derived extracellular vesicle; RCA, Ricinus communis agglutinin; SNA, Sambucus nigra ; sWGA, succinylated wheat germ agglutinin; UEA, Ulex europaeus agglutinin.

Article Snippet: Ricinus communis agglutinin 1, succinylated wheat germ agglutinin, Sambucus nigra , Maackia amurensis , concanavalin A, and Ulex europaeus agglutinin (all fluorescein-conjugated) were from Vector Laboratories.

Techniques: Comparison, Generated, Isolation, Flow Cytometry, Concentration Assay, Binding Assay, Fluorescence, Derivative Assay

Journal: bioRxiv

Article Title: Epithelial function of the circadian clock gene, Bmal1 , in regulating the mucosa

doi: 10.64898/2026.04.15.718752

Figure Lengend Snippet: ( a )Representative images of Epi Ctrl and Epi ΔBmal1 distal colon stained for UEA-1 (violet) and MALII (yellow) to label fucosylated and sialylated mucins, respectively. Shown are samples collected at ZT0 and ZT12, arrowheads indicate mucus thickness. (Scale bar = 20μm). ( b ) UEA-1+ mucus thickness in the distal colon is significantly higher at ZT12 in Epi Ctrl but lower at both times in Epi ΔBmal1 (**p≤0.01). ( c ) MALII+ mucus thickness in the distal colon is significantly higher at ZT12 in Epi Ctrl (*p≤ 0.05), with a significant increase of thickness in Epi ΔBmal1 at ZT0 (*p<0.05). Note, that Epi ΔBmal1 does not show a time-of-day dependent change in MALII. ( d ) UEA-1+ goblet cell area in distal colon crypts s significantly higher at ZT12 in Epi Ctrl (**p≤0.05). While not significantly different from controls, Epi ΔBmal1 do not show this time-of-day change in UEA-1+ mucus thickness. ( e ) MALII+ goblet cell area in distal colon crypts is significantly increased in Epi ΔBmal1 at ZT0 (*p≤0.05).

Article Snippet: Sections were incubated with biotinylated MAL-II (B-1265-1, Vector Laboratories; 3μg/mL in 1%BSA in PBS) overnight at 4°C, washed in PBS, then incubated with UEA-1 rhodamine (RL-1062-2, Vector Laboratories; 2μg/mL) and streptavidin-FITC (405201, BioLegend; 5μg/mL) in 1% BSA in PBS for 1 hr at room temperature in the dark.

Techniques: Staining

Journal: PLoS ONE

Article Title: Estradiol Increases Mucus Synthesis in Bronchial Epithelial Cells

doi: 10.1371/journal.pone.0100633

Figure Lengend Snippet: 10 −7 M estradiol (black bars) increased FUT-4, -5 and -6 mRNA expression compared with vehicle control (white bars). Data are expressed as A) absolute change in C t value normalized to HPRT1 and B) fold increase in mRNA expression over vehicle control normalized using HPRT1. Values shown are mean ± SEM of 4 different donors performed in 3 biological replicates. Total fucose sugar residues in the cytoplasmic protein fractions of ALI cultures were determined by lectin binding assays. Fucose residues were significantly increased with 10 −7 M estradiol using fucose binding lectins, C-D) AAA, E-F) LTA and G-H) UEA-1. D, F, and H are densitometric quantifications of C, E, and G, respectively. The intensity of all bands in each lane in detecting total fucose residues were quantified using the software program Image J and normalized to total protein loaded per lane in milligrams. Data is expressed as fold increase over vehicle control. Values shown are mean ± SEM of experiments performed with N = 4 donors. * P<0.05, ** P<0.01 compared against vehicle control. Non-parametric t-tests were used in all statistical analyses.

Article Snippet: Membranes were incubated with primary antibodies against ER-α, ER-β, NFATc1, MUC5AC, or (LTA-lotus tetragonolobus asparagus pea: H1601-1, AAA-anguilla anguilla lectin from fresh water eel: H4901-1, and UEA-1-ulex europaeus lectin from gorse: H2201-1) lectins (EY Laboratories Inc., San Mateo, CA) followed by incubation of secondary HRP-conjugated anti-rabbit, anti-mouse and beta-actin antibodies for 1 h at room temperature, and visualized using an enhanced chemiluminescence substrate (Thermo Scientific, Ontario, CAN) and a Chemigenius imaging system (Syngene, Cambridge, UK).

Techniques: Expressing, Control, Binding Assay, Software