APC BrdU Flow Kit

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	- Cells were incubated for 1h with bromodeoxyuridine.

Protocol tips
- Cells were incubated for 1h with bromodeoxyuridine.

Publication protocol

HMCLs were cultured for 4 days in 96-well flat-bottom microtiter plates in RPMI 1640 medium, 10% FCS and 2 ng/ml IL-6 (control medium) in the presence of various compounds. Cell growth was evaluated by quantifying intracellular ATP amount with a Cell Titer Glo Luminescent Assay (Promega, Madison, WI, USA) using a Centro LB 960 luminometer (Berthold Technologies, Bad Wildbad, Germany). Doxycycline (1 mg/ml) was added to induce shRECQ1 expression. The cell cycle was assessed using 49,6-diamidino-2-phenylindole (DAPI) staining (Sigma-Aldrich, Munich, Germany), incubation with bromodeoxyuridine (BrdU) for 1 h, and labeling with an anti-BrdU antibody (APC BrdU flow kit, BD Pharmingen, Le Pont De Claix, France) as described.41 Apoptotic cells were detected using phycoerythrin-conjugated Annexin V (PE-annexin V, BD Pharmingen). Apoptosis was measured using caspase-3/7 and caspase-9 luminescence Assay (Caspase Glo Assay (Promega, Madison, WI, USA)) according to the manufacturer’s protocol.

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Manufacturer protocol

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