Journal: British Journal of Cancer

Article Title: Prostaglandin E 2 stimulates Fas ligand expression via the EP1 receptor in colon cancer cells

doi: 10.1038/sj.bjc.6604490

Figure Lengend Snippet: Reduction in Jurkat T-cell viability following co-culture with colon cancer cells treated with PGE 2 . HT29 and HCA-7 cells were serum-deprived for 48 h, followed by treatment for 24 h with 0.1% DMSO, 0.5 or 1.0 μ M PGE 2 . Cells were fixed and then co-cultured with Jurkat T cells for 24 h. Jurkat T-cell viability was determined by resazurin reduction. To block FasL-mediated apoptosis, Jurkat T cells were also co-cultured with 1.0 μ M PGE 2 -treated colon cancer cells in the presence of 10 μ g ml −1 recombinant Fas:Fc blocking protein. Data denote mean±s.e.m. and are representative of two independent experiments. Viability is expressed as an index (% of control Jurkat T cells maintained in medium only).

Article Snippet: After 24 h, Jurkat cell viability was assessed by measuring the increase in fluorescence intensity associated with the cellular reduction of resazurin to resorufin, according to the manufacturer's instructions (R&D Systems).

Techniques: Co-Culture Assay, Cell Culture, Blocking Assay, Recombinant

Journal: Cells

Article Title: Anti-Inflammatory Extract from Soil Algae Chromochloris zofingiensis Targeting TNFR/NF-κB Signaling at Different Levels

doi: 10.3390/cells11091407

Figure Lengend Snippet: Inhibitory effects of NAE_2022C on TNFα- and UVB-induced inflammatory responses. ( A ) Brightfield micrographs of C. zofingiensis cultivated at standard growth conditions (stage 1; Upper) and after nitrogen deprivation stress (stage 2; Lower). Scale bar, 10 μm. ( B ) NAE_2022C inhibited TNFα- but not UVB-induced NF-κB responses. HaCaT cells were incubated with 100 nM DM, 750 nM WFA or 10 µg mL −1 NAE_2022C for 1 h and subsequently stimulated with 0.75 ng mL −1 TNFα or irradiated with 0.15 J/cm 2 UVB. The luciferase activity was measured in cell lysates 6 or 24 h post-stimulation, respectively (left y-axis). The cell viability was evaluated using a resazurin assay and shown as orange squares (right y-axis). Data are presented as mean ± SD ( n = 3). ( C ) The suppression of TNFα-induced pro-inflammatory mediators in reconstituted human epidermis (epiCS) by NAE_2022C. The release of cytokines and chemokines was determined in the medium after the treatment of epiCS with 0.75 ng mL −1 TNFα for 24 h (MCP1, RANTES and TNFα) and 48 h (CXCL10) using the Luminex Assay. Data represent the mean ± SD of at least three independent biological replicates. * p < 0.05 vs. samples treated with DMSO and TNFα.

Article Snippet: The cytotoxicity of algae extracts was evaluated using the Resazurin Cell Viability Kit (Cell Signaling Technology Europe B.V., Frankfurt am Main, Germany) and CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega GmbH, Walldorf, Austria) according to the manufacturers’ protocols.

Techniques: Incubation, Irradiation, Luciferase, Activity Assay, Resazurin Assay, Luminex