Journal: Respiratory Research

Article Title: Anti-inflammatory potential of PI3Kδ and JAK inhibitors in asthma patients

doi: 10.1186/s12931-016-0436-2

Figure Lengend Snippet: Effects of dexamethasone, PIK-294 and tofacinib on TCR-induced pSTAT5 activation in lymphocytes. Isolated blood T-cells from asthma patients ( n = 5) and healthy subjects ( n = 6) and BAL cells ( n = 4 asthma plus n = 3 healthy) were treated with 1000nM dexamethasone, PIK-294 or tofacinib for 1 h before TCR-stimulation for 4 h. Levels of phosphorylated STAT5 in CD3 + cells were quantified by flow cytometry. Data is presented as mean % pSTAT5 positive CD3 cells +/− standard deviation. Drug effects were assessed by 1-way ANOVA with a Dunnett’s multiple comparison test against the stimulated no drug control: * p < 0.05

Article Snippet: Cytotoxic effects of the drugs were assessed in TCR-stimulated PBMCs by Pierce LDH assay (Life Technologies, Paisley, UK) and propidium iodide flow cytometry assay (BD Bioscience, Oxford, UK); Additional file : Figure S1.

Techniques: Activation Assay, Isolation, Flow Cytometry, Standard Deviation

Journal: Respiratory Research

Article Title: Anti-inflammatory potential of PI3Kδ and JAK inhibitors in asthma patients

doi: 10.1186/s12931-016-0436-2

Figure Lengend Snippet: Expression of PI3Kδ in subepithelial and BAL T-cells from asthma patients and healthy subjects. PI3Kδ expression was examined in CD3 cells in subepithelial bronchial tissue and BAL by immunohistochemistry and flow cytometry, respectively. Representative images show PI3Kδ expression in bronchial tissue from healthy subjects ( a ) and asthma patients ( b ). Arrows and amplified images highlight example staining: Red for CD3 only; Blue for PI3Kδ only; Green for dual CD3 and PI3Kδ. Black scale bars represent 50 μm. Data is presented as ( c ) numbers of CD3 + PI3Kδ + cells per mm 2 subepithelium (Healthy n = 10; Asthma n = 9); ( d ) percentage of subepithelial CD3 cells expressing PI3Kδ (Healthy n = 8; Asthma n = 8); ( e ) percentage of BAL CD3 cells expressing PI3Kδ (Healthy n = 6; Asthma n = 9) and ( f ) PI3Kδ-dependent mean fluorescence intensity in dual labelled CD3 + PI3Kδ + BAL cells (Healthy n = 6; Asthma n = 9), with bar illustrating mean values. The percentage of subepithelial CD3 cells expressing PI3Kδ could not be calculated for n = 2 healthy and n = 1 asthma, due to lack of CD3 cells. Differences between healthy and asthma were assessed by T-test: * p < 0.05; ** p < 0.01

Article Snippet: Cytotoxic effects of the drugs were assessed in TCR-stimulated PBMCs by Pierce LDH assay (Life Technologies, Paisley, UK) and propidium iodide flow cytometry assay (BD Bioscience, Oxford, UK); Additional file : Figure S1.

Techniques: Expressing, Immunohistochemistry, Flow Cytometry, Amplification, Staining, Fluorescence

Journal: Neural Regeneration Research

Article Title: Yizhijiannao Granule and a combination of its effective monomers, icariin and Panax notoginseng saponins , inhibit early PC12 cell apoptosis induced by beta-amyloid (25–35) ☆

doi: 10.3969/j.issn.1673-5374.2012.24.001

Figure Lengend Snippet: PC12 apoptosis by flow cytometry. (A) Nerve growth factor group. (B) Model group. (C) Icariin + Panax notoginseng saponins (PNS) group. (D) PNS group. (E) Yizhijiannao Granule-containing serum group. (F) Icariin group. Propidium iodide (PI) is used to label cells in the middle and late stages of apoptosis and Annexin V is used to label early apoptotic cells. Annexin V-fluorescein isothiocyanate (FITC) + /PI cells are early apoptotic cells. The left superior quadrant represents necrotic cells; the left inferior quadrant represents normal cells; the right superior quadrant represents early apoptotic cells; and the inferior quadrant represents non-viable apoptotic cells. Early PC12 apoptosis was significantly reduced in the Yizhijiannao Granule-containing serum and icariin + PNS groups compared with the icariin or PNS groups.

Article Snippet: The early apoptosis rate of PC12 cells was detected using Annexin V-fluorescein isothiocyanate/propidium iodide staining flow cytometry (Becton Dickinson) according to the Annexin V-fluorescein isothiocyanate kit (Bender Med Systems, Vienna, Austria).

Techniques: Flow Cytometry

Journal: Cell Death & Disease

Article Title: 𝒟(−)lentiginosine-induced apoptosis involves the intrinsic pathway and is p53-independent

doi: 10.1038/cddis.2012.97

Figure Lengend Snippet: Susceptibility to apoptosis of U937Bcl-2 and U937pMEP cells. Both transfectants were treated with 𝒟(−)lentiginosine at the concentration of 10, 50, 100, 250, 500 and 1000 μ M for 18 h. The data on Y-axis are expressed as percentage hypodiploid nuclei, as detected by propidium iodide staining and flow cytometry analysis. As a positive control, etoposide at the concentration of 50 μ M was used. Data are presented as mean values±S.D. from three experiments performed in duplicate ( n =6). Asterisks (**) indicate highly significant differences between treated and untreated cells ( P <0.001)

Article Snippet: Apoptosis was assessed through flow cytometry analysis of isolated nuclei stained with propidium iodide, carried out on a Becton Dickinson FACS analyser, or by morphological analysis following staining with Hoechst chromatin dye, as previously described by some of us.,

Techniques: Concentration Assay, Staining, Flow Cytometry, Positive Control