annexin fitc 1 binding buffer  (R&D Systems)

Journal: Journal of Medicinal Chemistry

Article Title: Optimization of Antiproliferative Properties of Triimine Copper(II) Complexes

doi: 10.1021/acs.jmedchem.4c01806

Figure Lengend Snippet: Quantification of viable HCT116-DoxR tumor cells, in early apoptosis, late apoptosis, and necrosis marked with Annexin V-FITC and PI, was performed by flow cytometry after exposure to Cu1a and Cu1b complexes for 48 h at their IC 50 concentrations. Three controls (DMSO 0.1%, Cis, and Dox) were also included for comparison. The presented results are expressed as mean ± SEM obtained from at least two independent biological assays. To assess the statistical significance of these results compared to the control group treated with DMSO, the one-way ANOVA method was used (* p ≤ 0.05; ** p ≤ 0.005; *** p ≤ 0.0005; **** p ≤ 0.0001).

Article Snippet: Subsequently, the samples were incubated at room temperature (RT) with Alexa Fluor 488 annexin V and 100 μg/mL of propidium iodide (PI) in the dark for 15 min. After incubation, 1× Annexin V-FITC binding buffer was added and the samples were analyzed using an Attune Acoustic Focusing Flow Cytometer (Life Technologies, Carlsbad, USA).

Techniques: Flow Cytometry, Comparison, Control

Journal: Frontiers in Endocrinology

Article Title: Exploring lncRNAs associated with human pancreatic islet cell death induced by transfer of adoptive lymphocytes in a humanized mouse model

doi: 10.3389/fendo.2023.1244688

Figure Lengend Snippet: Characterization of islet-lncRNAs in islets cultured with glucose and with and without cytokines. (A) Changes in expression of islet-lncRNA, genes related to β-cell secretion (insulin, PCSK1), related to apoptosis (NFKβ1, CASP3, BAX, and BCL2), and related to ER stress (DDIT3 and HSPA5) in response to glucose stimulation. Data presented as mean ± SE. ***Paired t -test, p < 0.05. (B) Images of immunohistochemistry coupled with TUNEL assays on the islets cultured in vitro with cytokines (lower panel) and control (upper panel). DNA was stained as blue with DAPI, TUNEL + cells were detected as green, and insulin + cells/β cells were identified red. Apoptotic β cells are indicated as white arrows. (C) Percentage of apoptotic TUNEL + β cells in total β cells per islets cultured with and without cytokines from three donors (paired t -test, p < 0.05). (D) Determination of the effect of cytokines on apoptosis of islet cells in flow cytometry assay. Dot plots demonstrating separation of dissociated islet cells into early apoptotic cells in the lower right quadrant (Q3) (Annexin V +/ PI - ), necrotic cells in the upper right quadrant (Q2) (Annexin V + and PI + ), and late apoptotic cells in upper left quadrant (Q1) (Annexin V - and PI + ). Value in percentage of lower left quadrant (Q1) represents viable cells that were not stained with Annexin-V or PI. (E) Fold changes of lncRNAs in the cytokine-treated islets compared to that of control islets. Data presented as mean ± SE. *** Paired t -test, p < 0.05.

Article Snippet: Dissociated cells with media were filtered through a cell strainer (35 μm), washed twice with PBS, and resuspended in 1× Annexin V Binding Buffer from the FITC Annexin V Apoptosis Detection kit (BD Biosciences) in a concentration of 1.0 × 10 6 cells/100 μL.

Techniques: Cell Culture, Expressing, Immunohistochemistry, TUNEL Assay, In Vitro, Staining, Flow Cytometry

Journal: Frontiers in Endocrinology

Article Title: Exploring lncRNAs associated with human pancreatic islet cell death induced by transfer of adoptive lymphocytes in a humanized mouse model

doi: 10.3389/fendo.2023.1244688

Figure Lengend Snippet: Staining of exosomes and detection of lncRNA in exosomes in response to cytokines. (A) Images of exosomes precipitated from culture media of cytokine-treated islets (lower panel) and from control islets (upper panel). DNA was stained as blue with DAPI, Annexin-V was detected as green, and the phospholipid bilayer membrane of exosomes was stained as red. Scale bars represent 1 µm. (B) A hierarchically clustered heatmap of the relative expression of islet-lncRNAs in the exosomes precipitated from islets (from three donors) cultured with and without cytokines. Treatment numbers 1, 2, and 3 represent the islets from donors 1, 2, and 3, respectively.

Article Snippet: Dissociated cells with media were filtered through a cell strainer (35 μm), washed twice with PBS, and resuspended in 1× Annexin V Binding Buffer from the FITC Annexin V Apoptosis Detection kit (BD Biosciences) in a concentration of 1.0 × 10 6 cells/100 μL.

Techniques: Staining, Membrane, Expressing, Cell Culture

Journal: Nature Communications

Article Title: Proteomic characterization of gastric cancer response to chemotherapy and targeted therapy reveals potential therapeutic strategies

doi: 10.1038/s41467-022-33282-0

Figure Lengend Snippet: a Kaplan–Meier plots show significant differences between the sensitive group (S) and non-sensitive group (NS) in overall survival (OS) (upper) and progression-free survival (PFS) (bottom) in the HER2 subcohort (two-sided log rank test). b Boxplot showing the ERBB2 expression in HSG ( n = 32) and HNSG ( n = 37) (two-sided Wilcoxon rank-sum test) (upper). Boxplots show median (central line), upper and lower quartiles (box limits), 1.5× interquartile range (whiskers). Cox analysis (two-sided Cox test) of ERBB2 with OS (bottom). c Heatmap showing the abundance of differentially expressed proteins in HSG and HNSG. d Pathway alterations in HSG and HNSG (two-sided Fisher’s exact test). e The GSEA enrichment analysis of ECM in HER2 subcohort (Nominal P value, calculated as Phenotype-based permutation test). The survival analysis of ssGSEA ECM pathway score with OS in HER2 subcohort (two-sided log rank test). f Heatmap illustrating the abundance of ECM proteins in HER2 subcohort and five gastric cancer cells (two-sided Wilcoxon rank-sum test). g , h Left panel: Correlation of ECM pathway score ( g ) and PI3K-AKT pathway score ( h ) with its downstream pathway assessed by ssGSEA (two-sided Pearson’s correlation test). Right panel: Heatmap illustrating the protein abundance of PI3K-AKT pathway ( g ) and apoptosis ( h ) related proteins (two-sided Wilcoxon rank-sum test). i Dose-response curves of NCI-N87 cells after 72-h treatments with buparlisib (BUP), trastuzumab (TRA), and combination of trastuzumab and buparlisib with a ratio of 1:1. IC50, half-maximal inhibitory concentration. The comparison of IC50 values of different therapies (two-sided Student’s t test). Bars represent the mean of n = 3 independent experiments with error bars indicating SD. j Diagram showing the potential mechanism of resistance to anti-HER2 targeted therapy. k The decision-making strategy for the GC therapy. * P < 0.05 is considered statistically significant. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Source data are provided as a Source Data file.

Article Snippet: After two washes with cold PBS, cells were resuspended in 100 μL 1 × Binding Buffer (Annexin V-FITC/PI Apoptosis Detection Kit, YEASEN, Shanghai, China).

Techniques: Expressing, Quantitative Proteomics, Concentration Assay, Comparison