siGENOME Human BRCA1 (672) siRNA - SMARTpool

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	ES2 ovarian cancer cells were verified by short tandem repeat profiling by The Centre for Applied Genomics Genetic Analysis Facility (Toronto, ON, Canada). All cells were grown in Dulbecco’s modified Eagle’s medium/F-12 supplemented with 10% FBS, 100 U/mL penicillin, 100 mg streptomycin and fungizone (Gibco Cell Culture Products/Invitrogen). A549 cells are a commonly used cell line for studies on NF-kB signaling. Si GENOME SMARTpool BRCA1-targeting siRNA, and nontargeting siRNA were purchased from Dharmacon. Western blotting confirmed specific knockdown of BRCA1 (80%) relative to nontargeting siRNA control. To determine the impact of BRCA1 on NF-kB transcriptional activity, cells were plated at 100,000 cells/well in 24-well plates and transiently transfected with 50 pmol each of DAB2, or nontargeting siRNA 24 hours after plating using Lipofectamine LTX (Invitrogen) according to the manufacturer’s protocol.

Protocol tips

ES2 ovarian cancer cells were verified by short tandem repeat profiling by The Centre for Applied Genomics Genetic Analysis Facility (Toronto, ON, Canada). All cells were grown in Dulbecco’s modified Eagle’s medium/F-12 supplemented with 10% FBS, 100 U/mL penicillin, 100 mg streptomycin and fungizone (Gibco Cell Culture Products/Invitrogen). A549 cells are a commonly used cell line for studies on NF-kB signaling. Si GENOME SMARTpool BRCA1-targeting siRNA, and nontargeting siRNA were purchased from Dharmacon. Western blotting confirmed specific knockdown of BRCA1 (80%) relative to nontargeting siRNA control. To determine the impact of BRCA1 on NF-kB transcriptional activity, cells were plated at 100,000 cells/well in 24-well plates and transiently transfected with 50 pmol each of DAB2, or nontargeting siRNA 24 hours after plating using Lipofectamine LTX (Invitrogen) according to the manufacturer’s protocol.

Publication protocol

ES2 ovarian cancer cells were verified by short tandem repeat profiling by The Centre for Applied Genomics Genetic Analysis Facility (Toronto, ON, Canada). All cells were grown in Dulbecco’s modified Eagle’s medium/F-12 supplemented with 10% FBS, 100 U/mL penicillin, 100 mg streptomycin and fungizone (Gibco Cell Culture Products/Invitrogen). A549 cells are a commonly used cell line for studies on NF-kB signaling. Si GENOME SMARTpool BRCA1-targeting siRNA, and nontargeting siRNA were purchased from Dharmacon. Western blotting confirmed specific knockdown of BRCA1 (80%) relative to nontargeting siRNA control. To determine the impact of BRCA1 on NF-kB transcriptional activity, cells were plated at 100,000 cells/well in 24-well plates and transiently transfected with 50 pmol each of DAB2, or nontargeting siRNA 24 hours after plating using Lipofectamine LTX (Invitrogen) according to the manufacturer’s protocol.

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