Journal: Frontiers in Immunology

Article Title: Amelioration of Endotoxemia by a Synthetic Analog of Omega-3 Epoxyeicosanoids

doi: 10.3389/fimmu.2022.825171

Figure Lengend Snippet: (A) Polarization protocol to M1 macrophages by THP-1 cells. The histogram shows the MFI ratio of FITC for anti-human CD80 antibodies and the percentage of M1 macrophages. (B) The polarization protocol to M2 macrophages by U937 cells. The histogram shows the MFI ratio of Alexa Fluor 647 for anti-human CD209 antibodies and the percentage of M2 macrophages. **: < 0.0001, *: 0.0001 to 0.05. All experiments were triplicated.

Article Snippet: Phagocytosis was assessed using a Phagocytosis Assay Kit IgG-FITC (Cayman Chemical, Ann Arbor, USA).

Techniques:

Journal: International Journal of Nanomedicine

Article Title: In vitro apoptotic and DNA damaging potential of nanobarium oxide

doi: 10.2147/IJN.S95734

Figure Lengend Snippet: ( A – C ) Flow cytometirc analysis of Annexin V–FITC/PI stained cells. ( D ) Percentage of Annexin V–FITC/PI-positive cells in control and L929 cells exposed to BaONPs (150 μg/mL) for 24 and 48 hours. Notes: Each value represents the average ± SE of two experiments. Statistical differences with respect to the controls are shown (* P <0.05). Abbreviations: BaONPs, barium oxide nanoparticles; FITC, fluorescein isothiocyanate; PI, propidium iodide; SE, standard error.

Article Snippet: Apoptosis was detected by using the Annexin V–FITC (fluorescein isothiocyanate) kit (Cayman Chemicals, Ann Arbor, MI, USA) according to manufacturer’s information.

Techniques: Staining, Control

Journal:

Article Title: Helicobacter pylori Induces Apoptosis of Macrophages in Association with Alterations in the Mitochondrial Pathway

doi: 10.1128/IAI.72.5.2889-2898.2004

Figure Lengend Snippet: H. pylori induces apoptosis, as measured by annexin-V binding to externalized phosphatidylserine. RAW 264.7 cells were infected with H. pylori at an MOI of 50:1 for 24 h. Viable cells (annexin-V− PI−); nonviable, including late apoptotic or necrotic cells (annexin-V+ PI+ or annexin-V− PI+); and apoptotic cells (annexin-V+ PI−) were detected by the binding of Ann-V to externalized phospatidylserine in conjunction with PI, a dye excluded from viable cells. (A, B, and C) One FACS analysis representative of three individual experiments. (A) Eighty percent of uninfected RAW 264.7 cells were viable (annexin-V− PI−), while only 5% were apoptotic (annexin-V+ PI−). (B) One percent of RAW 264.7 cells treated with 1 μM staurosporine (positive control for apoptosis) were apoptotic (annexin-V+ PI−), and 98% of RAW 264.7 cells were nonviable (annexin-V+ PI+). (C) Sixteen percent of RAW 264.7 cells infected with H. pylori (60190) at an MOI of 50:1 for 24 h were apoptotic (annexin-V+ PI−), and 31% of the cells were nonviable (annexin-V+ PI+). FL1, flow cytometry channel 1 to detect PI stain; FL2, flow cytometry channel 2 to detect annexin-V. (D) Combined results of three separate FACS analyses depicting the mean levels of apoptotic cells (annexin-V+ PI−). Staurosporine-treated cells showed an increase in apoptosis over uninfected controls. RAW 264.7 cells infected with H. pylori (60190) at an MOI of 50:1 for 24 h showed an increase in apoptosis in comparison with uninfected controls (16.0% ± 3.7% versus 2.8% ± 0.6%; Tukey-Kramer test; *, P < 0.05).

Article Snippet: The binding of annexin V-fluorescein isothiocyanate (Ann-V) to externalized phosphatidylserine was used as a measurement of apoptotic RAW 264.7 cells with an Ann-V-propidium iodide (PI) apoptosis detection kit (BD Biosciences, Montreal, Quebec, Canada) according to the manufacturer's instructions.

Techniques: Binding Assay, Infection, Positive Control, Flow Cytometry, Staining

Journal:

Article Title: Helicobacter pylori Induces Apoptosis of Macrophages in Association with Alterations in the Mitochondrial Pathway

doi: 10.1128/IAI.72.5.2889-2898.2004

Figure Lengend Snippet: vacA and cagA are involved in H. pylori-induced apoptosis of RAW 264.7 cells. RAW 264.7 cells infected with H. pylori strain 60190 (24 h; MOI, 100:1) showed an increase in apoptosis (annexin-V+ PI−) in comparison with uninfected controls (13.8% ± 1.8% versus 2.4% ± 1.1%; ANOVA; **, P < 0.001) as assessed by Ann-V-PI binding. However, isogenic mutant strains deficient in either cagA (24 h; MOI, 100:1) or vacA (24 h; MOI, 100:1) showed a reduction in apoptosis (annexin-V+ PI−) compared to RAW 264.7 cells infected with the wild-type strain (7.6% ± 0.35% and 5.7% ± 1.5%, respectively, versus 13.8% ± 1.8%; ANOVA-Tukey-Kramer test; *, P < 0.002; n = 3).

Article Snippet: The binding of annexin V-fluorescein isothiocyanate (Ann-V) to externalized phosphatidylserine was used as a measurement of apoptotic RAW 264.7 cells with an Ann-V-propidium iodide (PI) apoptosis detection kit (BD Biosciences, Montreal, Quebec, Canada) according to the manufacturer's instructions.

Techniques: Infection, Binding Assay, Mutagenesis

Journal:

Article Title: Effects of zileuton and montelukast in mouse experimental spinal cord injury

doi: 10.1038/sj.bjp.0707577

Figure Lengend Snippet: Effect of zileuton and montelukast on apoptosis. At 24 h after SCI, spinal cord tissues obtained from SCI+vehicle-treated mice demonstrated a marked appearance of positive stain for annexin V FITC (a), an index of cells that are induced to undergo apoptosis. Some cells showed a positive intracellular staining to PI (a1), an index of cells in the late stage of apoptosis. On the contrary, spinal cord tissues section from mice treated with zileuton or montelukast demonstrate no cells in the earlier stage (b and c, respectively) of apoptosis and fewer cells (positive to PI) in the later stages of apoptosis (b1 and c1, respectively). (a2, b2, and c2) Staining combination of a and a1, b and b1, and c and c1, respectively. The figure is representative of at least three experiments performed on different experimental days. FITC, fluorescein isothiocyanate; PI, propidium iodide SCI, spinal cord injury.

Article Snippet: The binding of annexin-V fluorescein isothiocyanate (FITC) (Ann-V) to externalized phosphatidylserine was used as a measurement of apoptosis in spinal cord tissue section with an Ann-V-propidium iodide (PI) apoptosis detection kit (Santa Cruz, DBA, Milan, Italy) according to the manufacturer's instructions.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Gelsolin, an Actin-Binding Protein: Bioinformatic Analysis and Functional Significance in Urothelial Bladder Carcinoma

doi: 10.3390/ijms242115763

Figure Lengend Snippet: Effect of GSN knockdown on cell cycle and apoptosis. ( a ) Cell cycle distribution of T24 cells 72 h post-transfection with GSN-specific siRNA (siGSN#1) or non-targeted control siRNA (siNC#1). ( b ) Bar chart represents the percentage of cells in G1, S and G2/M phases of the cell cycle following GSN knockdown. The cell number in each phase was calculated using FlowJo software ( https://www.flowjo.com/ ). ( c , d ) Annexin V-FITC flow cytometry shows a significant increase in the apoptosis of T24 cells after 72 h of transfection with GSN siRNA (siGSN#1) compared with nontargeted control treated siRNA group (siNC#1). ( e ) Caspase-3/7 activity of T24 cells transfected with siGSN#1 was increased compared with the caspase-3/7 activity of cells transfected with control siNC#1 as determined by an Apo-ONE Homogeneous Caspase-3/7 assay. All data are represented as the mean ± SD of three independent experiments. *** p < 0.001.

Article Snippet: siRNA-transfected cells undergoing early/late apoptosis were analyzed with annexin V-FITC and PI staining using annexin an V-FITC assay kit (Cayman Chemical, Ann Arbor, MI, USA).

Techniques: Transfection, Software, Flow Cytometry, Activity Assay