Journal: Heliyon

Article Title: Influence of iron binding in the structural stability and cellular internalization of bovine lactoferrin

doi: 10.1016/j.heliyon.2021.e08087

Figure Lengend Snippet: Internalization of bLf in Vero cells. BLf in its apo (A) or holo (B) forms were conjugated to FITC and synchronously incubated with Vero cells for 0, 30 or 60 min. BLf internalization was observed by laser-scanning confocal fluorescence microscopy. The selected groups of cells are representative of their respective open fields. White bars: 20 μm.

Article Snippet: Apo- and holo-bLf were incubated with FITC (Molecular Probes, Eugene, OR, USA) at a molar ratio of 1:10 in basic phosphate buffer (2.5% Na 2 HPO 4 .7H 2 O and 0.082% NaH 2 PO 4 at pH 8.0) for 1 h at 4 °C.

Techniques: Incubation, Fluorescence, Microscopy

Journal: BMC Gastroenterology

Article Title: Glucagon-like peptides 1 and 2 and vasoactive intestinal peptide are neuroprotective on cultured and mast cell co-cultured rat myenteric neurons

doi: 10.1186/1471-230x-12-30

Figure Lengend Snippet: Figure 1 Immunocytochemical staining of myenteric neurons cultured in absence (A) or presence (B-F) of peritoneal mast cells. A. Neurons grown for 6 days and stained with PGP9.5. The cultured neurons survive well; they group into ganglion-like structures and grow a prominent arborizing network of nerve terminals. B. Neurons co-cultured with mast cells (arrows) stained with antibodies against PGP9.5 and histamine, respectively. In the neuronal cultures to which mast cells have been added the number of neurons markedly decreases and the terminal network is disintegrated. C and D. show mast cells grown in co-culture and stained with histamine antibodies. The mast cells are well settled within the cultures and they contain a high number of cytoplasmic granules. C. illustrates a cultured mast cell with numerous granules in the central, perinuclear, portion of the cell. D. shows a mast cell exhibiting piece meal degranulation. E and F. illustrate the morphological relationship of cultured neurons and mast cells. E. shows mast cells (arrowheads) in close contact with nerve terminals from nearby myenteric neurons (arrows). F. a mast cell and a nerve cell body in close proximity. In order to perceive the morphological arrangement of mast cells and neurons both PGP9.5- and histamine- immunoreactive cells are visualised by FITC conjugated antibodies. Neurons and mast cells are distinguished by their morphological characteristics. Bar in A 100 μm represents also B, bar in C 40 μm represents also D, bar in E 50 μm represents also F.

Article Snippet: For visualization, the slides were exposed to fluorescein isothiocyanate (FITC)- or Texas Red- conjugated goat anti-rabbit IgG antiserum (Jackson Immunoresearch Laboratories, West Grove, PA, USA; dilution 1:100) or FITC conjugated goat anti-mouse IgG antiserum (Jackson Immunoresearch Laboratories; dilution 1:100) for 1 h, mounted in phosphate buffer:glycerol 1:1 and analyzed using a fluorescence microscope with appropriate filter settings.

Techniques: Staining, Cell Culture, Co-Culture Assay

Journal: The European respiratory journal

Article Title: Classical protein kinase C and its hypoxic stimulus-induced translocation in the cat and rat carotid body.

doi: 10.1034/j.1399-3003.2000.016003459.x

Figure Lengend Snippet: Fig. 1. ± Confocal fluorescence microscopy of double immunolabelling of a section of a normoxic cat carotid body tissue stained with mouse anti- protein kinase C (PKC)c and rabbit anti-tyrosine hydroxylase (TH). The localization of PKCc is visualized in green with fluorescein isothiocyanate labelled antimouse immunoglobulin G (IgG) (A), whereas that of TH in red with Texas red-labelled antirabbit IgG (B). An overlapping image is shown in C; the yellow spots indicate colocalization of the two fluorescent probes. The near-perfect colocalization identifies the PKCc as being in the chemoreceptor cell. (Internal scale bars=10 mm.)

Article Snippet: For immunofluorescence staining, sections were incubated with fluorescein isothiocyanate (FITC)-labelled antimouse immunoglobulin G (IgG; dilution 1:200; Jackson Immunoresearch West Grove, PA, USA) for 1 h. To confirm the chemoreceptor cell localization of the PKC antigens, other sections were double-labelled by simultaneous incubation with a rabbit polyclonal antibody directed against rat tyrosine hydroxylase (TH; dilution 1:500; Chemicon International, Inc., Temecula, CA, USA) and visualization with Texas red-labelled antirabbit IgG (dilution 1:200; Jackson Immunoresearch).

Techniques: Fluorescence, Microscopy, Staining

Journal: Viruses

Article Title: Baculovirus PTP2 Functions as a Pro-Apoptotic Protein

doi: 10.3390/v10040181

Figure Lengend Snippet: Transient expression of the Spodoptera exigua protein tyrosine phosphatase 2 (SePTP2) induced mild apoptosis in Sf21 cells. ( A ) Overview of the expression cassette in the pIB-DEST vector: egfp and the gene of interest were separated by the foot and mouth disease virus (FMDV) 2A region and expression of the cassette was driven by the OpIE2 promoter; ( B ) EGFP expression in Sf21 cells at 48 h post-transfection (hpt) with pIB-EGFP + ActD (ActD + EGFP), pIB-EGFP (EGFP), pIB-SePTP2 (SePTP2), or pIB-SePTP2mut (SePTP2mut), respectively. Apoptotic bodies are indicated by white circles and arrows, scale bar = 200 µm; ( C ) caspase activity levels in Sf21 cells transfected with pIB-EGFP + ActD (ActD + EGFP), pIB-SePTP2 (SePTP2), pIB-SePTP2mut (SePTP2mut), pIB-AcPTP (AcPTP), pIB-EGFP (EGFP), pIB-DESTmod (DESTmod) and mock. Caspase 3/7 activity in transfected Sf21 cells was measured as relative luminescent units (RLUs) at 48 hpt, relative to a blank containing cell medium. Error bars represent the standard error of the mean. Treatment groups marked with a different letter (a, b, c or d) are significantly different .

Article Snippet: Next, 5 µL of Annexin V-EGFP (Annexin V-EGFP Apoptosis Detection Kit, BioVision, Milpitas, CA, USA) was added to stain any apoptotic cells, while Hoechst dye was added at a concentration of 10 µM to stain the nuclei of all cells.

Techniques: Expressing, Plasmid Preparation, Transfection, Activity Assay

Journal: Viruses

Article Title: Baculovirus PTP2 Functions as a Pro-Apoptotic Protein

doi: 10.3390/v10040181

Figure Lengend Snippet: Caspase inhibitor carbobenzoxy-valyl-alanyl-aspartyl-(O-methyl)-fluoromethylketone (Z-VAD-FMK) blocked induction of apoptosis in Sf21 cells expressing SePTP2. EGFP expression in Sf21 cells at 48 h post transfection with pIB-SePTP2 ( A ) or with pIB-SePTP2 + Z-VAD-FMK ( B ). Apoptotic bodies are indicated by white circles and arrows, scale bar = 200 µm.

Article Snippet: Next, 5 µL of Annexin V-EGFP (Annexin V-EGFP Apoptosis Detection Kit, BioVision, Milpitas, CA, USA) was added to stain any apoptotic cells, while Hoechst dye was added at a concentration of 10 µM to stain the nuclei of all cells.

Techniques: Expressing, Transfection

Journal: Viruses

Article Title: Baculovirus PTP2 Functions as a Pro-Apoptotic Protein

doi: 10.3390/v10040181

Figure Lengend Snippet: Fluorescence microscopy analysis of cells in S. exigua hemocytes at 48 h post transfection. Cells were stained with Hoechst (left panel) and Annexin V-EGFP (middle panel). Cells were obtained from larvae infected with wild-type (WT) Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV), ∆ ptp2 SeMNPV or no virus (mock), scale bar = 200 µm.

Article Snippet: Next, 5 µL of Annexin V-EGFP (Annexin V-EGFP Apoptosis Detection Kit, BioVision, Milpitas, CA, USA) was added to stain any apoptotic cells, while Hoechst dye was added at a concentration of 10 µM to stain the nuclei of all cells.

Techniques: Fluorescence, Microscopy, Transfection, Staining, Infection