Journal: Langmuir : the ACS journal of surfaces and colloids

Article Title: Single Molecule Spectroscopy Studies of Acid-Base Chemical Gradients Using Nile Red as a Probe of Local Surface Acidity.

doi: 10.1021/acs.langmuir.1c02059

Figure Lengend Snippet: Figure 3. Solution phase ensemble fluorescence from Nile Red in mixtures of ethyl acetate (EthAc) and acetic acid (HOAc). (a) Fluorescence spectra obtained as a function of HOAc in EthAc using a conventional fluorimeter. (b) Ensemble, solution phase E values obtained from Nile Red (10 nM) in a series of HOAc/EthAc mixtures, showing an abrupt change in its emission spectrum with increasing acid concentration. The data in (b) were recorded on the same microscope used for single molecule experiments.

Article Snippet: Dye-doped aminosilane gradient films were imaged on a wide-field epifluorescence microscope that has been described previously.36 Briefly, this microscope is built upon a Nikon TiE inverted light microscope that employs the Nikon Perfect Focus stabilization system.

Techniques: Fluorescence, Concentration Assay, Microscopy

Journal: Journal of leukocyte biology

Article Title: Angiopoietin chemotactic activities on neutrophils are regulated by PI-3K activation.

doi: 10.1189/jlb.0906580

Figure Lengend Snippet: Fig. 1. Detection of Tie2 expression on neutrophil cell surface by FACScan analysis. Neutrophils were incubated with mouse monoclonal Tie2 IgG1 (@Tie2; 1, 5, or 10 g/ml) or with control mouse isotypic IgG1 (Control IgG1; 1, 5, or 10 g/ml). Neutrophils were then incubated with a secondary, goat antimouse, FITC-conjugated IgG (1:100). Flow cytometric analysis (10,000 events) was performed using a FACScan. Representative analysis of three independent experiments.

Article Snippet: In another set of experiments, cells were pretreated for 15 min with a blocking goat polyclonal antihuman Tie2 (anti-hTie2) IgG (R&D Systems) or isotypic goat IgG (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) before cell migration toward angiopoietins (1 nM).

Techniques: Expressing, Incubation, Control

Journal: Journal of leukocyte biology

Article Title: Angiopoietin chemotactic activities on neutrophils are regulated by PI-3K activation.

doi: 10.1189/jlb.0906580

Figure Lengend Snippet: Fig. 2. Ang1 and Ang2 mediate neutrophil migration through Tie2 activation. Neutrophils were added to the upper wells of a modified Boyden chamber apparatus, and the lower wells were filled with RPMI PBS, Ang1, or Ang2 (0.1–10 nM; A). In another series of experiments, neutrophils were pretreated with a blocking goat anti-hTie2 IgG (@Tie2; 5 or 10 g/ml) or control goat isotypic IgG (IgG; 5 or 10 g/ml) for 15 min prior to the addition of neutrophils in the upper wells of a modified Boyden chamber apparatus in which the lower wells were filled with RPMI PBS, Ang1, or Ang2 (1 nM; B). Treatment with CXCL8/IL-8 (25 nM; lower wells) was used as a positive control. Migrating cells were fixed, stained, and counted under light microscopy at 400 original magnification. Values are means SEM of at least three independent experi- ments. *, P 0.05; **, P 0.01; ***, P 0.001, compared with PBS. ††, P 0.01; †††, P 0.001, as compared with corresponding agonist.

Article Snippet: In another set of experiments, cells were pretreated for 15 min with a blocking goat polyclonal antihuman Tie2 (anti-hTie2) IgG (R&D Systems) or isotypic goat IgG (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) before cell migration toward angiopoietins (1 nM).

Techniques: Migration, Activation Assay, Blocking Assay, Control, Positive Control, Staining, Light Microscopy

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Diagrams of the in vitro NVU system and the double cell co-culture systems. Astrocytes were seeded in the outer side of the matching Transwell inserts after neurons were seeded into a 24-well culture plate and cultured for 5–7 days. After 2 days of co-culture, endothelial cells were seeded in the inner side of the matching well inserts. The NVU triple cell system and the double cell co-culture systems were successfully established after 3 days of culture. NE: Neuron; AS: astrocyte; BM: brain microvascular endothelial cell; NO: no cell; NVU: neurovascular unit.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: In Vitro, Co-Culture Assay, Cell Culture

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Morphological and immunocytochemical characteristics of the hippocampal NVU. (A) Hippocampal neurons, astrocytes and microvascular endothelial cells under an inverted light microscope. (B) Neurons are positive for NSE (arrows), astrocytes express GFAP (arrows), and brain microvascular endothelial cells are positive for CD31 (arrows). Scale bars: 100 μm. NVU: Neurovascular unit; NSE: neuron-specific enolase; GFAP: glial fibrillary acidic protein.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Light Microscopy

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Expression of occludin protein in the hippocampal NVU triple cell co-culture system and in the AS + BM co-culture system, with or without glucose (150 mM) and corticosterone (200 μM), assessed by western blot assay. Results are shown as the mean ± SD ( n = 3; one-way analysis of variance followed by Dunnett’s post hoc test). * P < 0.05, ** P < 0.01, vs . NVU group; # P < 0.05, vs . NVU model group (NVU + glucose + corticosterone). Western blot assay was performed in triplicate. GAPDH was used as loading control. The numbers on the right are the molecular weights of the proteins (kDa). AS: Astrocyte; BM: brain microvascular endothelial cell; NVU: neurovascular unit; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Expressing, Co-Culture Assay, Western Blot, Control

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Transendothelial electrical resistance (A) and leakage liquid height (B) in the hippocampal NVU triple cell co-culture system and in the AS + BM co-culture system, with or without glucose (150 mM) and corticosterone (200 μM), as detected by Millipore resistor and vernier caliper. Quantitative results are shown as the mean ± SD ( n = 3; one-way analysis of variance followed by Dunnett’s post hoc test). * P < 0.05, ** P < 0.01, vs . NVU group; ## P < 0.01, vs . NVU model group (NVU + glucose + corticosterone); $ P < 0.05, $$ P < 0.01, vs . AS + BM group. Experiments were performed in triplicate. NE: Neuron; AS: astrocyte; BM: brain microvascular endothelial cell; NVU: neurovascular unit; TEER: transendothelial electrical resistance.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Co-Culture Assay

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Expression of CX43 and GFAP proteins in the hippocampal NVU triple cell co-culture system and in the AS + BM co-culture system, with or without glucose (150 mM) and corticosterone (200 μM), as detected by western blot assay. Results are shown as the mean ± SD ( n = 3; one-way analysis of variance followed by Dunnett’s post hoc test). * P < 0.05, ** P < 0.01, vs . NVU group; # P < 0.05, vs . NVU model group (NVU + glucose + corticosterone). Western blot assay was performed in triplicate. GAPDH was used as loading control. Numbers on the right indicate the molecular weights of the proteins (kDa). NE: Neuron; AS: astrocyte; BM: brain microvascular endothelial cell; NVU: neurovascular unit; CX43: connexin 43; GFAP: glial fibrillary acidic protein.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Expressing, Co-Culture Assay, Western Blot, Control

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Intracellular and supernatant levels of 5-HT in the hippocampal NVU triple cell co-culture system and in the AS + BM co-culture system, with or without glucose (150 mM) and corticosterone (200 μM), as measured by enzyme-linked immunosorbent assay. Results are shown as the mean ± SD ( n = 3; one-way analysis of variance followed by Dunnett’s post hoc test). ** P < 0.01, vs . NVU group; ## P < 0.01, vs . NVU model group (NVU + glucose + corticosterone); $ P < 0.05, vs . AS + BM group. Experiments were performed in triplicate. NE: Neuron; AS: astrocyte; BM: brain microvascular endothelial cell; NVU: neurovascular unit; F&M: fluoxetine and metformin; 5-HT: 5-hydroxytryptamine.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Apoptosis in the hippocampal NVU triple cell co-culture system and in the AS + BM co-culture system, with or without glucose (150 mM) and corticosterone (200 μM), as detected by flow cytometry. Results are shown as the mean ± SD ( n = 3; one-way analysis of variance followed by Dunnett’s post hoc test). ** P < 0.01, vs . NVU group; # P < 0.05, vs . NVU model group (NVU + G & P, NVU + glucose + corticosterone). All apoptotic cells were in the Q2 and Q4 quadrants. Early apoptotic cells were in Q4, while late apoptotic cells were in Q2. Experiments were performed in triplicate. NE: Neuron; AS: astrocyte; NVU: neurovascular unit; G&P: glucose and corticosterone; F&M: fluoxetine and metformin.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Co-Culture Assay, Flow Cytometry

Journal: Neural Regeneration Research

Article Title: Structural and functional damage to the hippocampal neurovascular unit in diabetes-related depression

doi: 10.4103/1673-5374.244794

Figure Lengend Snippet: Apoptosis in the hippocampal NVU triple cell co-culture system and in the AS + BM co-culture system, with or without glucose (150 mM) and corticosterone (200 μM), as detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling staining. Arrows point to apoptotic cells. NE: Neuron; AS: astrocyte; NVU: neurovascular unit; G & P: glucose and corticosterone; F & M: fluoxetine and metformin. Scale bars: 100 μm.

Article Snippet: Astrocytes and brain microvascular endothelial cells were labeled with rabbit anti-glial fibrillary acidic protein (GFAP) (1:100; Boster) and rabbit anti-PECAM-1/CD31 (1:100; Boster) antibodies.

Techniques: Co-Culture Assay, End Labeling, Staining