Journal: Cellular and Molecular Life Sciences

Article Title: Sialic acid blockade in dendritic cells enhances CD8 + T cell responses by facilitating high-avidity interactions

doi: 10.1007/s00018-021-04027-x

Figure Lengend Snippet: Reduced sialylation favors clustering between BMDCs and CD8 + OT-I T cells Ac 5 3F ax Neu5Ac and control-treated BMDCs were pulsed with OVA peptide or medium. For cell identification during flow cytometry acquisition, BMDCs were labeled with CellTrace Violet dye, and CD8 + OT-I T cells were labeled with CellTrance Far Red dye. After 45 min of incubation at 37 °C, cluster formation was assessed by flow cytometry. Representative dot plots show BMDCs–CD8 + OT-I T cell clusters (left) and quantification is shown as bar diagram (right). Percentages are shown as mean values ± SD of two biological replicates and experiments were performed three times

Article Snippet: CD8 + OT-I T cells were fluorescently labeled with CellTrace Far Red dye (Thermo Fisher Scientific) according to manufacturer’s protocol and 1–3*10 5 cells were flushed into the z-Movi Chip.

Techniques: Flow Cytometry, Labeling, Incubation

Journal: CNS Neuroscience & Therapeutics

Article Title: Endothelial Progenitor Cells Contribute to Neointima Formation in Rabbit Elastase‐Induced Aneurysm after Flow Diverter Treatment

doi: 10.1111/cns.12086

Figure Lengend Snippet: BM‐derived EPCs are tracked in neointima. In the early 2 weeks after autologous transfusion, EPCs with double fluorescence are observed in the subendothelium space (A–B) and around the stent struts(C–D). In the late 2 weeks after autologous transfusion, EPCs with double fluorescence are observed on the surface of neointima (E–H). Double‐positive cells were identified by showing yellow emission fluorescence for CFSE and blue emission fluorescence for Hoechst 33,342 concurrently.

Article Snippet: EPCs at the second to fourth passage were harvested at day 14, 17, and 20 of culture, double labeled with the DNA‐specific fluorochrome Hoechst 33,342 (1 μg/mL, Sigma, USA) for one hour and the intracellular fluorescent dye CFSE (carboxyfluorescein diacetate succinimidyl ester, 10 μmoL/mL, Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 15 min at 37°C, suspended in 10 mL saline, and autologously transfused via the margin ear vein.

Techniques: Derivative Assay, Fluorescence

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Enhancement of Endometrial Receptivity by Cnidium officinale through Expressing LIF and Integrins

doi: 10.1155/2019/7560631

Figure Lengend Snippet: The effect of CoM on cytotoxicity and the adhesion of JAr cell to Ishikawa cells. (a) Ishikawa cells (1.5 × 10 6 cells) were seeded and incubated with CoM at indicated concentration (up to 500 μ g/ml) for 24 h. The cell viability was measured by MTT assay. The data were expressed as percentage of controls and are shown as mean ± SEM of three independent experiments. (b) Ishikawa cells were treated with CoM (50 μ g/mL) for 48 h. CMFDA-labeled JAr cells were added onto the CoM-treated Ishikawa cells. The attached JAr cells were fixed, pictured using fluorescent microscopy, and manually counted. Data were calculated as mean ± SEM of three independent experiments. The data were expressed as fold of controls. ∗∗ p < 0.01 for comparison between two groups (magnification ×50; scale bar = 5 μ m).

Article Snippet: To make cell monolayers, Ishikawa cells (1.5 × 10 6 cells) were seeded into 6-well plates and incubated with CoM treatment for 48 h. The JAr cells (2 × 10 6 cells) were labeled with 5-chloromethylfluorescein diacetate (CMFDA) fluorescence dye (CellTracker Green; Life Technologies, Carlsbad, CA) for 15 min. To check adhesion between Ishikawa and JAr cells, labeled JAr cells were added to the Ishikawa cell monolayer.

Techniques: Incubation, Concentration Assay, MTT Assay, Labeling, Microscopy

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Enhancement of Endometrial Receptivity by Cnidium officinale through Expressing LIF and Integrins

doi: 10.1155/2019/7560631

Figure Lengend Snippet: Effect of antagonist for LIFR and neutralization for integrins α V, β 3, and β 5 on the adhesion of JAr cells to CoM-stimulated Ishikawa cells. (a) Ishikawa cells were treated with or without LIF antagonist (hLA) (50 ng/mL) for 1 h and then CoM (50 μ g/ml) was added to hLA-pretreated Ishikawa cells for 48 h. (b) Ishikawa cells (1.5 × 10 6 cells) were cultured in 6-well plates and treated with or without CoM (50 μ g/ml) for 48 h, and the cells were then incubated in the presence of integrin α V, β 3, β 5, and IgG antibodies for 2 h. CMFDA-labeled JAr cells were added onto Ishikawa cell monolayer. The attached cells were fixed and pictured using fluorescent microscopy. Data were calculated as mean ± SEM of three independent experiments and expressed as fold of controls. ∗∗∗ p < 0.001 compared to each group (magnification ×50; scale bar = 5 μ m).

Article Snippet: To make cell monolayers, Ishikawa cells (1.5 × 10 6 cells) were seeded into 6-well plates and incubated with CoM treatment for 48 h. The JAr cells (2 × 10 6 cells) were labeled with 5-chloromethylfluorescein diacetate (CMFDA) fluorescence dye (CellTracker Green; Life Technologies, Carlsbad, CA) for 15 min. To check adhesion between Ishikawa and JAr cells, labeled JAr cells were added to the Ishikawa cell monolayer.

Techniques: Neutralization, Cell Culture, Incubation, Labeling, Microscopy