Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Role of leukotriene A 4 hydrolase aminopeptidase in the pathogenesis of emphysema 1

doi: 10.4049/jimmunol.1400452

Figure Lengend Snippet: Assessment of the LTA4H protein expression in 129sv wild type mice post exposure to cigarette smoke over 5 months. Panel A: The levels of LTA4H protein in the whole lung BALF assessed by ELISA. Panel B: The levels of LTA4H protein in the whole lung protein soup assessed by ELISA. Panel C: Flow cytometry of the whole lung single cell suspension. Air = mice exposed to ambient air for 20 weeks. SM = mice exposed to cigarette smoke for 20 weeks. Panel D: Immunohistochemistry with antibody specific to murine LTA4H protein in lung tissues from mice exposed to cigarette smoke for 20 weeks (63× oil magnification). Positive signals appear brown and counterstained with blue. Arrowheads show nuclei with positive counterstain (blue) indicating absence of LTA4H protein. Arrows show nuclei with positive stain (grown) indicating presence of LTA4H protein. Gray bars in Panels A & B = ambient-air exposed mice with ages 26 – 28 weeks. * represents analysis by ANOVA, ** by Bonferroni subgroup comparison, and *** by nonparametric t-Test. N = 5 per group.

Article Snippet: Next, all CD45 high cells were gated into Ly6G high and CD11b high cells (neutrophils) as previously described. ( 3 ) For the purpose of detecting cells containing the LTA 4 H in their intra-cellular space, cells were permeabilized with flow cytometry permeabilization buffer (R&D) after stained with CD45 antibody.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Immunohistochemistry, Staining

Journal: Frontiers in Pharmacology

Article Title: Estrogens Modulate Somatostatin Receptors Expression and Synergize With the Somatostatin Analog Pasireotide in Prostate Cells

doi: 10.3389/fphar.2019.00028

Figure Lengend Snippet: Effect of estradiol on cell cycle. Flow cytometry analysis of propidium iodide-labeled EPN (A) or CPEC (B) cells was done, as described in Methods. Quiescent cells were left under basal conditions or treated with estradiol (20 nM; E 2 ) for the indicated times. Histograms represent the % counts/FL2 areas of labeled cells in the different cell cycle phases after 24 or 48 h of hormonal treatment. Means and SEMs are shown. n represents the number of experiments. ∗ p < 0,05 for each experimental point vs. the corresponding untreated control.

Article Snippet: Permeabilization was performed by incubating 10 6 cells in 0,5 ml of flow cytometry analysis (FACS) permeabilizing solution, which contains RNAses and propidium iodide (Beckton Dickinson).

Techniques: Flow Cytometry, Labeling, Control

Journal: Frontiers in Pharmacology

Article Title: Estrogens Modulate Somatostatin Receptors Expression and Synergize With the Somatostatin Analog Pasireotide in Prostate Cells

doi: 10.3389/fphar.2019.00028

Figure Lengend Snippet: Effect of both estradiol and pasireotide on cell cycle. Flow cytometry analysis of propidium iodide-labelled EPN (A) or CPEC (B) was done, as described in Methods. In panel (A,B) , quiescent cells were left under untreated or treated for 24 h with 0,1 μM pasireotide, in the absence or presence of 20 nM estradiol. Histograms in panel (A,B) represent the % counts/FL2 areas of labeled cells in the different cell cycle phases after the indicated treatments. Means and SEMs are shown. n represents the number of experiments. ∗ p < 0,05; ∗∗ p < 0,01. In panel (C) , quiescent EPN (left section) or CPEC (right section) cells were left untreated or treated for 24 h with 0,1 μM pasireotide, in the absence or presence of 20 nM estradiol. Lysate proteins were separated by SDS-PAGE, transferred to PDVF membrane and filters were then analyzed by Western blot, using the anti-activated caspase 3 or anti-tubulin antibodies.

Article Snippet: Permeabilization was performed by incubating 10 6 cells in 0,5 ml of flow cytometry analysis (FACS) permeabilizing solution, which contains RNAses and propidium iodide (Beckton Dickinson).

Techniques: Flow Cytometry, Labeling, SDS Page, Membrane, Western Blot

Journal:

Article Title: Female X-chromosome mosaicism for gp91phox expression diversifies leukocyte responses during endotoxemia

doi: 10.1097/CCM.0b013e3181eb9ed6

Figure Lengend Snippet: Lipopolysaccharide (LPS)-induced changes in bone marrow (BM) and blood cell composition in WT, gp91phox-deficient and mosaic animals. Twenty-four hours after LPS or saline controls, BM cells and blood were collected and processed for flow cytometry analyses, as described in Materials and Methods. Graphs A and B depict BM myeloid cell content identified by CD45/CD11b double positive (A) and B-cell content identified by CD45/CD19 double positive staining (B). Graphs C–F depict the numbers of circulating CD11b+ (mostly neutrophils) and different T-cell (CD4+ and CD8+) and B-cell (CD19+) subsets as indicated. Cell numbers were calculated from percent distribution of cells and total white blood cell yield. Statistically significant difference (p < .05): *compared with control within the same genotype; #compared with WT or heterozygous controls. Mean ± SEM, n = 7–8 animals in each group.

Article Snippet: Flourochrome conjugated antibodies, assay diluents, lysing and permeabilizing flow cytometry solutions and kits were purchased from BD Biosiences (San Jose, CA) and BD Pharmingen (San Diego, CA).

Techniques: Flow Cytometry, Staining