Journal: BMC veterinary research

Article Title: Therapeutic effect of long-interval repeated subcutaneous administration of canine amniotic membrane-derived mesenchymal stem cells in atopic dermatitis mouse model.

doi: 10.1186/s12917-025-04554-w

Figure Lengend Snippet: Fig. 5 Expression profiles of CD markers in T cells determined by flow cytometry analysis after stimulation of canine PBMCs with conA. (A) Representa tive FACs image of CTL, conA, conA + MSCs and ConA + Pime. PBMCs were treated with concanavalin A (conA; 5 μm/ml), conA combined with MSCs (1 × 105cells), conA combined with pimecrolimus (Pime; 100 ng/ml) until Day 8. The markers used were CCR7 (CD197) and CD45ra. Three independent analyses were conducted. (B) Representative FACs image of CTL, conA, conA + MSCs and ConA + Pime. PBMCs were treated with concanavalin A (conA; 5 μm/ml), conA combined with MSCs (1 × 105cells), conA combined with pimecrolimus (Pime; 100 ng/ml) until Day 8. The markers used were CD62L and CD45ra. Three independent analyses were conducted. (C) Quantification of CCR7-negative and CD45ra-positive cells through flow cytometry analysis in triplicate. mean ± SD; ** p < 0.01, *** p < 0.001. (D) Quantification of CD62L-negative and CD45ra-positive cells through flow cytometry analysis in triplicate. mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The cells were stained with CCR7 (C-C chemokine receptor type 7; clone: 3D12, 1:100 dilution, BD science, USA, Catalog number: 552176), cluster of differentiation 45ra (CD45ra; clone: HI100, 1:500 dilution, BD science, Catalog number: 550855), and CD62L (L-selectin; clone: FMC46, 1:100 dilution, Novus bio, Catalog number: NB100-65388) for two hours at 4 °C under blocking light.

Techniques: Expressing, Flow Cytometry

Journal: Pathogens

Article Title: The Involvement of Neutrophil in the Immune Dysfunction Associated with BVDV Infection

doi: 10.3390/pathogens12050737

Figure Lengend Snippet: The effect of BVDV-infected MDM supernatants (296C, 1373, and 28508-5 sup) on neutrophil surface marker expression: ( A ) CD18, ( B ) and L-selectin. Sup: supernatant *: p < 0.05 (>95% confidence). The result is the average of at least 3 replicates ( n = 3).

Article Snippet: Fifty (50) μL of primary antibodies of anti-CD14 antibody (clone M-M9; VMRD Inc., Pullman, WA, USA), anti-CD18 antibody (clone BAQ30A; Kingfisher Biotechnology, St Paul, MN, USA), or anti-L selectin antibody (clone FMC46; Novus Biological, Littleton, CO, USA) were added to respective wells in triplicates.

Techniques: Infection, Marker, Expressing

Journal: Thrombosis and haemostasis

Article Title: IL-9 Promotes the Development of Deep Venous Thrombosis by Facilitating Platelet Function.

doi: 10.1055/s-0038-1673614

Figure Lengend Snippet: Fig. 3 P-selectin surface levels on mouse platelets. (A) Representative pictures of CD62P expression in CTRL, interleukin (IL)-9, adenosine diphosphate (ADP), ADP þ IL-9 and AG490 þ ADP þ IL-9 groups. CD61 was used to identify platelets. Cells in the Q2 zone were platelets with P-selectin expression. (B) The results of statistical analysis on the ratio of CD62P in CTRL, IL-9, ADP, ADP þ IL-9 and AG490 þ ADP þ IL-9 groups, n ¼ 3. p < 0.05 versus CTRL (untreated), ADP and IL-9 þ ADP.

Article Snippet: The membranes were blocked with Tris-buffered saline with Tween 20 (TBST) (10 mM Tris-base, 100 mM NaCl and 0.01% Tween 20) containing 5% fat-free dry milk for 2 hours at room temperature, washed three times with TBST and incubated at 4°C for 24 hours with various primary antibodies, namely, IL-9 receptor (IL-9R) antibody (Santa Cruz Biotechnology, United States, 1:200), phosphor-JAK2 (CST,United States, 1:1,000), JAK2 (Abcam, United States, 1:1,000), phosphor-signal transducer and activator of transcription 3 (STAT3) (Abcam, 1:50,000), STAT3 (Abcam, 1:50,000), phosphor-AKT (CST, 1:1,000), AKT (CST, 1:1,000), IL-9R antibody (Santa, Europe, 1:200) and CD62P antibody (Novus, United States, 1:500).

Techniques: Expressing

Journal: Analytical Chemistry

Article Title: Nanoscopic Characterization of Cell Migration under Flow Using Optical and Electron Microscopy

doi: 10.1021/acs.analchem.2c04222

Figure Lengend Snippet: Characterization of the endothelium-like CHO-E monolayer. (a) Bright-field images of cultured CHO-E cells in the fluidic chamber. The cells were seeded over a glass-bottom fibronectin-coated surface. (b) Time-lapse experiment of CHO-E cell culture confluence. Total surface coverage is shown in percentage. (c) Viability of CHO-E cells before and after applying 2 dyne cm –2 shear stress for 10 min. (d) Bright-field and fluorescence images of fixed and stained CHO-E cells. E-selectin surface receptors were immunolabeled using primary antibodies and secondary AF-647 conjugated antibodies. (e) Box plots showing the number of E-selectin expressed on the surface of CHO-E and CHO-K cells. Each spot represents mean number of E-selectin in each cell. (f) Box plots showing the number of E-selectin at different locations on each CHO-E cell. Each spot represents the number of E-selectin in a different location on the same CHO-E cell. Data on total 10 CHO-E cells are displayed. Error bars represent ± standard deviation (SD) determined by n = 3 independent experiments in panels (b) and (c).

Article Snippet: A commercially available microfluidic chamber, μ-slide VI 0.5 glass bottom (3.8 mm width and 0.54 mm height, ibidi GmbH) was coated by a recombinant homodimeric human (rh) E-selectin (SELE, human protein, recombinant hlgG-Fc, His TAG, Sino Biological) to conduct the cell-rolling assay and fluorescence imaging experiments of KG1a cells on the recombinant E-selectin.

Techniques: Cell Culture, Fluorescence, Staining, Immunolabeling, Standard Deviation

Journal: Analytical Chemistry

Article Title: Nanoscopic Characterization of Cell Migration under Flow Using Optical and Electron Microscopy

doi: 10.1021/acs.analchem.2c04222

Figure Lengend Snippet: Rolling behavior of KG1a cells on CHO-E monolayer. (a) Time-lapse bright-field images of single KG1a cells rolling over a CHO-E monolayer at a shear stress of 2 dyne cm –2 . (b) Frame-to-frame single-cell velocity over 70 s time period. (c) Number of adhered then rolled KG1a cells over CHO-E and CHO-K monolayers in the presence or absence of EDTA. (d) Effect of the blocking of active binding sites on E-selectin on the time-averaged rolling velocity of KG1a cells on the CHO-E monolayer. The active binding sites were blocked by two different blocking antibodies, BB2G and H18-7. (e) Time-averaged rolling velocity of KG1a cells over CHO-E monolayer and rh E-selectin deposited on a glass surface with different wall shear stress values. Error bars represent ±SD determined by n = 3 independent experiments in panels (c)–(e).

Article Snippet: A commercially available microfluidic chamber, μ-slide VI 0.5 glass bottom (3.8 mm width and 0.54 mm height, ibidi GmbH) was coated by a recombinant homodimeric human (rh) E-selectin (SELE, human protein, recombinant hlgG-Fc, His TAG, Sino Biological) to conduct the cell-rolling assay and fluorescence imaging experiments of KG1a cells on the recombinant E-selectin.

Techniques: Blocking Assay, Binding Assay

Journal: Analytical Chemistry

Article Title: Nanoscopic Characterization of Cell Migration under Flow Using Optical and Electron Microscopy

doi: 10.1021/acs.analchem.2c04222

Figure Lengend Snippet: Scanning electron microscopy (SEM) characterization of nanoscopic morphology of KG1a cells during the cell rolling over the CHO-E monolayer. SEM images of KG1a cells deposited on the (a) coverslip and (b) CHO-E monolayer. (c) SEM image of KG1a cells rolled on a rh E-selectin-coated surface (0.2 μg mL –1 ) at a shear stress of 1 dyne cm –2 . (d) Enlarged view of the area highlighted by the white box in panel (c). Tethers formed during the cell rolling are highlighted by yellow boxes. Elongation of the KG1a cells is highlighted by ellipses. (e) SEM image of KG1a cells rolled on the CHO-E monolayer at a shear stress of 2 dyne cm –2 . KG1a cells are highlighted by circles. (f) Enlarged view of the area highlighted by the box in panel (e). Tethers formed during the cell rolling are highlighted by arrowheads. Tethering points are highlighted by red squares. (g) Box plots showing the length (top) and width (bottom) of the tethers and slings captured by SEM measurements.

Article Snippet: A commercially available microfluidic chamber, μ-slide VI 0.5 glass bottom (3.8 mm width and 0.54 mm height, ibidi GmbH) was coated by a recombinant homodimeric human (rh) E-selectin (SELE, human protein, recombinant hlgG-Fc, His TAG, Sino Biological) to conduct the cell-rolling assay and fluorescence imaging experiments of KG1a cells on the recombinant E-selectin.

Techniques: Electron Microscopy