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Transfection procedures

BG Benjamin Gottschalk
ZK Zhanat Koshenov
MW Markus Waldeck-Weiermair
SR Snježana Radulović
FO Furkan E. Oflaz
MH Martin Hirtl
OB Olaf A. Bachkoenig
GL Gerd Leitinger
RM Roland Malli
WG Wolfgang F. Graier
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HeLa cells were grown under standard culture conditions until 50% confluence was reached, transfected in DMEM (without FCS and antibiotics) with 1.5 µg/ml plasmids or 100 nM siRNA using 2.5 µg/ml TransFast™ transfection reagent (Promega, Madison, WI, USA). After 24 h, the medium was replaced with DMEM containing 10% FCS and 1% penicillin/streptomycin and kept for a further 24 h prior to experiments. The specific siRNAs (Microsynth, Balgach, Switzerland) used in this study are listed in Supplementary Table 1. Alternatively, plasmid transfection was done using PolyJet™ In Vitro DNA Transfection Reagent (SL100688, SignaGen® Laboratories, Frederick, MD, USA).

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