Journal: Stem Cell Reports

Article Title: Selective elimination of pluripotent stem cells by PIKfyve specific inhibitors

doi: 10.1016/j.stemcr.2021.12.013

Figure Lengend Snippet: WX8 induced programmed cell death selectively only in human pluripotent stem cells (A) Fibroblasts and ECCs were seeded at densities such that cells cultured in the absence of WX8 were not confluent at the end of the experiment. Cells were treated with various concentrations of WX8 for 4 days, stained with annexin-V and propidium iodide (PI) without prior permeabilization using an Annexin V-FITC Apoptosis Detection Kit, and then analyzed in a Becton Dickinson LSR-II using FACS Diva software. (B) SEM is indicated for average of three independent assays. Cells cultured in the presence of vehicle (0 μM WX8) were defined as 100% viable cells (Q3 • ▪) and 0% dead cells (Q1+Q2 ◯ □) so that only changes due to WX8 were represented. Time 0 h was plotted as 0.1 h to generate a logarithmic scale. Similar results were obtained with ESCs and iPSCs ( Figure S5 ).

Article Snippet: Fluorescein isothiocyanate (FITC) and propidium iodide (PI) fluorescence were detected with BD LSR-II using FACS Diva software (Becton Dickinson).

Techniques: Cell Culture, Staining, Software

Journal: Oncotarget

Article Title: Sensitization of glycoengineered interferon-β1a-resistant cancer cells by cFLIP inhibition for enhanced anti-cancer therapy

doi: 10.18632/oncotarget.14573

Figure Lengend Snippet: OVCAR-3, MCF-7, HeLa, or TOV-21G cells were treated with or without various concentrations of R27T. A . The cells were incubated for various times, and cell viability was measured using the WTS assay. Data are presented as the means ± SD of three independent experiments (*** P < 0.001 compared to the untreated group). B . Seventy-two hours after treatment, live cells were incubated with calcein-AM and analyzed by microscopy for detection of fluorescence. Scale bar = 250 μm. C . Apoptosis activity was quantified by bivariate flow cytometry of cells stained with Annexin V/PI at 72 h post-treatment.

Article Snippet: R27T-induced apoptosis was measured using a fluorescence isothiocyanate (FITC) Annexin V/PI kit (BD Bioscience, San Jose, CA, USA).

Techniques: Incubation, Microscopy, Fluorescence, Activity Assay, Flow Cytometry, Staining

Journal: Oncotarget

Article Title: Sensitization of glycoengineered interferon-β1a-resistant cancer cells by cFLIP inhibition for enhanced anti-cancer therapy

doi: 10.18632/oncotarget.14573

Figure Lengend Snippet: A-C . HeLa cells were transfected with 10 nM of scrambled or cFLIP siRNA for 24 h, and then treated with R27T (100 ng/mL) for an additional 48 h. Cell viability was analyzed by WTS assays (A) or live-cell staining (B), and apoptosis was assessed by Annexin V/PI staining (C). D . HeLa cells transfected with control, cFLIPL, cFLIPS, or cFLIPL + S vector were treated with TBB (10 μM) or R27T (100 ng/mL) or both for 48 h. Cell viability was analyzed by WTS assays. E-F . HeLa cells treated solely with TBB (10 μM) or R27T (100 ng/mL) or both for 48 h were stained with calcein-AM (E), and apoptosis was assessed by Annexin V/PI staining (F). Error bars indicate the SD of experiments performed in triplicate (*** P < 0.001 compared to the R27T-treated group). Scale bar = 250 μm.

Article Snippet: R27T-induced apoptosis was measured using a fluorescence isothiocyanate (FITC) Annexin V/PI kit (BD Bioscience, San Jose, CA, USA).

Techniques: Transfection, Staining, Plasmid Preparation