Journal: Cell Death & Disease

Article Title: Flavonol and imidazole derivatives block HPV16 E6 activities and reactivate apoptotic pathways in HPV + cells

doi: 10.1038/cddis.2015.391

Figure Lengend Snippet: 3-(1H-indol-1yl)propan-1-amine ( 1 ), three benzimidazole derivatives ( 2 – 4 ), and spinacine ( 5 ) inhibit protein–protein interactions. Five compounds at the indicated concentrations (1.4 μ M–3.2 mM) were tested using our bead-based screening assay for their ability to inhibit three different protein/protein interactions: ( a ) GST-E6/His-caspase 8; ( b ) GST-E6/HisE6AP; and ( c ) GST-caspase 8/His-caspase 8. Binding in the presence of 1.4 μ M of the test compound was set at 100%. Experiments were performed in triplicate, and error bars indicate the S.D.

Article Snippet: Caspase 3/7 activity was measured using the flourogenic substrate CellTitier-Glo for caspase 3/7 activity (Promega, Fitchburg, WI, USA) following the manufacturer's instructions.

Techniques: Protein-Protein interactions, Screening Assay, Binding Assay

Journal: Cell Death & Disease

Article Title: Flavonol and imidazole derivatives block HPV16 E6 activities and reactivate apoptotic pathways in HPV + cells

doi: 10.1038/cddis.2015.391

Figure Lengend Snippet: EC 50 ( μ M) values of tested small molecules for the indicated protein–protein interactions

Article Snippet: Caspase 3/7 activity was measured using the flourogenic substrate CellTitier-Glo for caspase 3/7 activity (Promega, Fitchburg, WI, USA) following the manufacturer's instructions.

Techniques: Protein-Protein interactions, Inhibition

Journal: Cell Death & Disease

Article Title: Flavonol and imidazole derivatives block HPV16 E6 activities and reactivate apoptotic pathways in HPV + cells

doi: 10.1038/cddis.2015.391

Figure Lengend Snippet: Myricetin and spinacine ( 5 ) increase caspase 3/7 activity in SiHa cells following treatment with TRAIL and chemotherapy drugs. SiHa cells (2 × 10 4 cells per well) were seeded into a 96-well plate and allowed to incubate overnight and then pretreated with 100 μ M of myricetin or 50 μ M of spinacine for 4 h. ( a ) TRAIL (100 ng/ml), along with cycloheximide (5 μ g/ml), ( b ) cisplatin (50 μ M), or ( c ) doxorubicin (2 μ M) were added respectively. Caspase 3/7 activity was measured after 0, 1.5, 3, and 6 h using the CellTiter-Glo assay. Activity at 0 h of treatment was set at 100% for each group. Experiments were performed in triplicate, and error bars indicate the S.D.

Article Snippet: Caspase 3/7 activity was measured using the flourogenic substrate CellTitier-Glo for caspase 3/7 activity (Promega, Fitchburg, WI, USA) following the manufacturer's instructions.

Techniques: Activity Assay, Glo Assay

Journal: Cell Death & Disease

Article Title: Flavonol and imidazole derivatives block HPV16 E6 activities and reactivate apoptotic pathways in HPV + cells

doi: 10.1038/cddis.2015.391

Figure Lengend Snippet: Treatment with myricetin and spinacine ( 5 ) increases cellular levels of caspase 8 and p53. ( a ) SiHa cells (1 × 10 6 per well) were seeded into a six-well plate and allowed to incubate overnight. The indicated concentrations of myricetin and spinacine were added, and then cells were incubated for 24 h. Cells were then washed in 1 × PBS and then harvested, and the resulting level of caspase 8 was measured by immunoblot. ( b ) SiHa and C33A cells (1 × 10 6 per well) were seeded into a six-well plate and allowed to incubate overnight. In all, 200 μ M of myricetin and 100 μ M spinacine were added together with 4 μ g/ml mitomycin C and incubated for 24 h. The resulting level of p53 was measured by enzyme-linked immunosorbent assay, and the level of p53 found in untreated cells was set at 100%. Experiments were performed in triplicate, and error bars indicate the S.D.

Article Snippet: Caspase 3/7 activity was measured using the flourogenic substrate CellTitier-Glo for caspase 3/7 activity (Promega, Fitchburg, WI, USA) following the manufacturer's instructions.

Techniques: Incubation, Western Blot, Enzyme-linked Immunosorbent Assay