Journal: bioRxiv

Article Title: Crosstalk in skin: Loss of desmoglein 1 in keratinocytes inhibits BRAF V600E -induced cellular senescence in human melanocytes

doi: 10.1101/2023.02.16.528886

Figure Lengend Snippet: Human primary melanocytes were transduced with WT BRAF or BRAF V600E lentivirus, control or Dsg1-deficient CM were added 24 h after transduction and cells continued to be cultured in CM before they were harvested. A , Relative cell numbers over time. B , EdU flow cytometry assay to determine cell proliferation. The figures showed a clear separation of proliferating cells which have incorporated EdU in control and WT BRAF melanocytes but not in BRAF V600E melanocytes. C , Quantification of EdU flow cytometry assay. D , Annexin V/dead cell apoptosis flow cytometry assay was performed to detect cell death. E , Quantification of cell death by flow cytometry assay. F , Senescence-associated β-galactosidase activity was detected by flow cytometry assay. Right shift of the peak indicated the increase in senescence. G , Quantification of senescence flow cytometry assay. H , Expression of the indicated proteins was determined by Western blot. Results were reported as mean ± SD (n=3), statistical analysis was performed using two-way ANOVA with multiple comparisons (*, P < 0.05; ***, P < 0.001).

Article Snippet: Following the same treatment as described above, a CellEvent Senescence Green Flow Cytometry Assay Kit (Thermo Fisher Scientific, C10841) was used per the manufacturer’s instructions.

Techniques: Transduction, Cell Culture, Flow Cytometry, Activity Assay, Expressing, Western Blot

Journal: iScience

Article Title: Resistance to spindle inhibitors in glioblastoma depends on STAT3 and therapy induced senescence

doi: 10.1016/j.isci.2024.111311

Figure Lengend Snippet:

Article Snippet: 72 h later cells were washed once with 1XPBS, fixed with 4% paraformaldehyde for 10 min, washed with PBS containing 1% BSA, and incubated with fluorescent β-galactosidase staining solution (CellEvent Senescence Green Detection Kit, cat# C10850, ThermoFisher Scientific) at 37 0 C in a dry incubator without CO 2 for 2 h. Cells were washed with 1XPBS and stained for F-actin and DAPI.

Techniques: Plasmid Preparation, Virus, Recombinant, Membrane, Protease Inhibitor, Western Blot, Lysis, Transfection, Bicinchoninic Acid Protein Assay, Isolation, Staining, Flow Cytometry, Software

Journal: Cell host & microbe

Article Title: Mitochondria-derived vesicles deliver antimicrobial reactive oxygen species to control phagosome-localized Staphylococcus aureus

doi: 10.1016/j.chom.2018.10.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cell death and Caspase-3/7 were measured by flow cytometry using SYTOX green dead cell stain and CellEvant Caspase-3/7 green flow cytometry assay kit according to manufacturer’s protocol (Thermo Fisher).

Techniques: Mutagenesis, Virus, Transformation Assay, Binding Assay, Control, Molecular Weight, Flow Cytometry, shRNA, Reverse Transcription, Transfection, Software, Imaging

Journal: bioRxiv

Article Title: Label-free ghost cytometry for manufacturing of cell therapy products

doi: 10.1101/2023.07.09.545884

Figure Lengend Snippet: (A) Caspase-3/7 activity and SytoxAADvanced scatter plot in which we gated live, apoptotic, and dead cells. (B) An SVM score histogram for the LF-GC-based classification of the live cells and dead/apoptotic cells in PBMCs (a.u. = arbitrary units). The ROC-AUC for this classification was 0.975.

Article Snippet: After thawing by immediately placing them into a 37°C water bath and transferring them into a pre-warmed RPMI1640 with 10% FBS and 1% Penicillin-Streptomycin, cells were stained with either the FITC Annexin V Apoptosis Detection Kit (BioLegend) or CellEvent™ Caspase-3/7 Green Flow Cytometry Assay Kit (Thermo Fisher Scientific) to label apoptotic cells (Caspase-3/7 activity-positive and SytoxAADvanced-negative) and dead cells (Caspase-3/7 activity-positive and SytoxAADvanced-positive) according to manufacturer instructions and analyzed using LF-GC.

Techniques: Activity Assay