ON-TARGETplus Human ARL2 (402) siRNA - Individual

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	Cells were grown in six-well culture dishes to 70–90% confluence. Plasmids (2 µg of pcDNA3.1 or pcDNA3.1 carrying indicated inserts and Mito-GFP; 2∶1 ratio) were co-transfected using LipofectAMINE and Plus reagents (Invitrogen) according to the manufacturer's instructions. siRNAs (25 nM) were transfected into HeLa cells using Dharmafect transfection reagent #1 (Dharmacon, Lafayette, CO) following the company's instructions. For some experiments, transfections were performed sequentially first with plasmid DNA followed 5 hours later by transfection of the ARL2 siRNA, to minimize cell toxicity evident from co-transfections. We consistently observed at least 70% transfection efficiency, using expression of fluorescence-tagged proteins to score

Protocol tips

Cells were grown in six-well culture dishes to 70–90% confluence. Plasmids (2 µg of pcDNA3.1 or pcDNA3.1 carrying indicated inserts and Mito-GFP; 2∶1 ratio) were co-transfected using LipofectAMINE and Plus reagents (Invitrogen) according to the manufacturer's instructions. siRNAs (25 nM) were transfected into HeLa cells using Dharmafect transfection reagent #1 (Dharmacon, Lafayette, CO) following the company's instructions. For some experiments, transfections were performed sequentially first with plasmid DNA followed 5 hours later by transfection of the ARL2 siRNA, to minimize cell toxicity evident from co-transfections. We consistently observed at least 70% transfection efficiency, using expression of fluorescence-tagged proteins to score

Publication protocol

Cells were grown in six-well culture dishes to 70–90% confluence. Plasmids (2 µg of pcDNA3.1 or pcDNA3.1 carrying indicated inserts and Mito-GFP; 2∶1 ratio) were co-transfected using LipofectAMINE and Plus reagents (Invitrogen) according to the manufacturer's instructions. siRNAs (25 nM) were transfected into HeLa cells using Dharmafect transfection reagent #1 (Dharmacon, Lafayette, CO) following the company's instructions. For some experiments, transfections were performed sequentially first with plasmid DNA followed 5 hours later by transfection of the ARL2 siRNA, to minimize cell toxicity evident from co-transfections. We consistently observed at least 70% transfection efficiency, using expression of fluorescence-tagged proteins to score

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