Journal: eLife

Article Title: Linker histone H1.2 and H1.4 affect the neutrophil lineage determination

doi: 10.7554/eLife.52563

Figure Lengend Snippet: ( a–c ) Analysis of circulating neutrophils and eosinophils in wild type (wt) and H1.2/H1.4-double deficient mice (H1.2/H1.4 -/-). p Values, derived from unpaired two-tailed T tests, are indicated. ( a ) Flow cytometry analysis of neutrophils (Gr1 positive, CD115 negative) in whole blood of adult age-matched animals (wt n = 15, H1.2/H1.4 - / - n = 14), other immune cell types are shown in . ( b ) Flow cytometry analysis of absolute amounts of neutrophils (Ly-6G positive cells) in whole blood (n = 4). ( c ) Flow cytometry analysis of amounts of eosinophils (SiglecF positive) in whole blood (n = 4). H1.2/H1.4 - / - animals show enhanced eosinophil numbers in circulation. d-f Lineage-negative hematopoietic stem cells were sorted from murine bone marrow and cultured for 6 days in the presence of various cytokines allowing differentiation into several immune cell lineages. Depicted are CD11b positive cells ( d ) and the fraction of neutrophils ( e ) or eosinophils ( f ) within CD11b-positive cells. Each dot represents an independent experiment consisting of two mice per genotype, the mean of the two wild type animals was set to one and the mean of the two H1.2/H1.4 - / - animals is depicted relative to wt. ( a–f ) Error bars are SEM.

Article Snippet: Recombinant murine cytokines (IL-3, IL-5, IL-9, GM-CSF, SCF, G-CSF) were from Peprotech.

Techniques: Derivative Assay, Two Tailed Test, Flow Cytometry, Cell Culture

Journal: eLife

Article Title: Linker histone H1.2 and H1.4 affect the neutrophil lineage determination

doi: 10.7554/eLife.52563

Figure Lengend Snippet: a, b mRNA expression of the chemokine CXCL1 ( a ) and the inflammatory cytokine IL6 ( b ) in homeostatic (left panel) and casein-injected (right panel) animals. H1.2/H1.4-deficient bone marrow cells transcribe more CXCL1 and less IL6 . ( c–e ) protein levels of the indicated cytokines and chemokines in the serum of homeostatic and casein-injected mice. The enhanced expression of CXCL1 is also seen in serum ( c ). Casein injection leads to an upregulation of IL-17 ( d ) and G-CSF ( e ) and the upregulation of IL-17 is stronger in H1.2/H1.4-deficient animals. Depicted is the mean -/+ SD, each data point represents one mouse. p values are indicated and derived from an unpaired, two-tailed t test.

Article Snippet: Recombinant murine cytokines (IL-3, IL-5, IL-9, GM-CSF, SCF, G-CSF) were from Peprotech.

Techniques: Expressing, Injection, Derivative Assay, Two Tailed Test

Journal: Theranostics

Article Title: Preliminary Therapy Evaluation of 225 Ac-DOTA-c(RGDyK) Demonstrates that Cerenkov Radiation Derived from 225 Ac Daughter Decay Can Be Detected by Optical Imaging for In Vivo Tumor Visualization

doi: 10.7150/thno.14338

Figure Lengend Snippet: In vivo blockade demonstrates specific targeting of α v β 3 integrin. α v β 3 + tumors demonstrate significantly more uptake of 225 Ac-DOTA-c(RGDyK) when compared to α v β 3 + tumors in animals receiving the radiotracer and excess c(RGDyK) as blockade.

Article Snippet: Recombinant human α v β 3 integrin was purchased from R&D Systems, Inc. (Minneapolis, MN).

Techniques: In Vivo

Journal:

Article Title: Interleukin-5 in growth and differentiation of blood eosinophil progenitors in asthma: effect of glucocorticoids

doi: 10.1038/sj.bjp.0704389

Figure Lengend Snippet: Proportion of CD34+ cells expressing intracellular IL-5 or surface IL-5Rα. (A) Representative flow cytometry plots of CD34+ cells expressing IL-5 or IL-5Rα from an asthma patient compared to a normal subject. (B) Both IL-5 and IL-5Rα expressions were significantly increased in patients with asthma (closed bars, n=8) compared to normal subjects (open bars, n=8). **P<0.01 compared with normal subjects. Data are mean±s.e.mean.

Article Snippet: In separate experimental groups, NANT cells were cultured in the presence or absence of exogenous human recombinant IL-5 (100 ng ml −1 ; R & D Systems, Minneapolis, MN, U.S.A.), or anti-IL-5Rα monoclonal neutralizing antibody (2.5 μg ml −1 ; Catalogue number AF-253-NA, R & D Systems, Minneapolis, MN, U.S.A.), or dexamethasone (10 −6 M ).

Techniques: Expressing, Flow Cytometry

Journal:

Article Title: Interleukin-5 in growth and differentiation of blood eosinophil progenitors in asthma: effect of glucocorticoids

doi: 10.1038/sj.bjp.0704389

Figure Lengend Snippet: Effect of IL-5Rα antibody, contaminating T cells and anti-sense IL-5 oligonucleotides on colony numbers after 14 days culture of progenitor cells. (A) Effect of IL-5Rα receptor blocking antibody (anti-IL-5R) in normal subjects (n=10) or patients with asthma (n=10). Anti-IL5R antibody inhibited colony numbers in asthmatics. **P<0.01 compared with normal subjects; #P<0.01 compared with corresponding control. (B) Colony numbers from non-adherent non-T cells from asthmatics subjects (n=14) compared to those obtained from non-adherent cells containing T cells from the same donors. (C) There were no significant changes in progenitor cells from normal subjects (n=8); in patients with asthma (n=6), transfection with IL-5 anti-sense oligonucleotides (OGN) 6 h before culture inhibited colony formation. **P<0.01 compared to those without transfection. Data are mean±s.e.mean.

Article Snippet: In separate experimental groups, NANT cells were cultured in the presence or absence of exogenous human recombinant IL-5 (100 ng ml −1 ; R & D Systems, Minneapolis, MN, U.S.A.), or anti-IL-5Rα monoclonal neutralizing antibody (2.5 μg ml −1 ; Catalogue number AF-253-NA, R & D Systems, Minneapolis, MN, U.S.A.), or dexamethasone (10 −6 M ).

Techniques: Blocking Assay, Control, Transfection

Journal:

Article Title: Interleukin-5 in growth and differentiation of blood eosinophil progenitors in asthma: effect of glucocorticoids

doi: 10.1038/sj.bjp.0704389

Figure Lengend Snippet: IL-5 positive (IL-5+) and IL-5 receptor positive (IL-5Rα+) cells (%) in non-adherent and non-T cell preparation after culture for 14 days

Article Snippet: In separate experimental groups, NANT cells were cultured in the presence or absence of exogenous human recombinant IL-5 (100 ng ml −1 ; R & D Systems, Minneapolis, MN, U.S.A.), or anti-IL-5Rα monoclonal neutralizing antibody (2.5 μg ml −1 ; Catalogue number AF-253-NA, R & D Systems, Minneapolis, MN, U.S.A.), or dexamethasone (10 −6 M ).

Techniques:

Journal:

Article Title: Interleukin-5 in growth and differentiation of blood eosinophil progenitors in asthma: effect of glucocorticoids

doi: 10.1038/sj.bjp.0704389

Figure Lengend Snippet: Effect of dexamethasone on expression of IL-5 and IL-5Rα in progenitor cells. Proportion of cells with intracellular IL-5 positive (A) and with surface expression of IL-5Rα (B) as detected by flow cytometry in non-adherent non-T cell preparation after 16 h pre-treatment with or without dexamethasone (10−6 M, Dex; +Dex or Time control) from patients with asthma (Asthma, n=7) and normal subjects (Normal, n=4). Dexamethasone significantly reduced IL-5 progenitor cells without affecting IL-5Rα+ progenitor cells in asthmatic patients. #P<0.01 compared with normal subjects; *P<0.05 compared with corresponding time control (n=4). Data are mean±s.e.mean.

Article Snippet: In separate experimental groups, NANT cells were cultured in the presence or absence of exogenous human recombinant IL-5 (100 ng ml −1 ; R & D Systems, Minneapolis, MN, U.S.A.), or anti-IL-5Rα monoclonal neutralizing antibody (2.5 μg ml −1 ; Catalogue number AF-253-NA, R & D Systems, Minneapolis, MN, U.S.A.), or dexamethasone (10 −6 M ).

Techniques: Expressing, Flow Cytometry, Control