celecoxib

Protocol tips

Upstream tips	Protocol tips	Downstream tips
FDA approved Cox-2 inhibitor. It is used to induce autophagy and apoptosis in cells.	Treat cells with 30 μM of celecoxib for 72 hours

Upstream tips
FDA approved Cox-2 inhibitor. It is used to induce autophagy and apoptosis in cells.
Protocol tips
Treat cells with 30 μM of celecoxib for 72 hours

Publication protocol

Cells were treated with DMSO or celecoxib (8 and 30 μM) for 72 hours. In some cases, U87MG cells were pre-treated with PFT (25 μM) for 30 minutes prior to celecoxib treatment. For apoptosis assays, trypsinised cells were incubated with FITC-conjugated Annexin V (BD Biosciences) and propidium iodide (5 μg/ml). 10,000 events were analysed for apoptosis by FACS with CellQuest Pro software. For autophagy assays, celecoxib-treated cells were stained with acridine orange (1 μg/ml, Sigma) for 15 mins at 37°C. Trypsinised cells were re-suspended in phenol red-free growth media (Gibco BRL) and 10,000 events were analysed by FACS with CellQuest Pro software. Acridine orange-stained cells grown on cover slips were viewed under a Laser Scanning Microscope and images captured using software LSM510.

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