Smartpool sirna oligos

siRNA transfection of U2Os was carried out using Lipofectamine^™ RNAiMAX transfection Reagent. Conditions used for RNA transfection in these cells were as described previously [11 (link)] [12 (link)]. Cells were transfected with SMARTpool siRNA oligos against human proteins, the results were then confirmed with single siRNA oligos (Dharmacon).
RAW264.7 cells were transfected with 100 pmol of SMARTpool siRNA oligos against mouse DCNL5, DCNL1, DCNL4 or a non-targeting control (Dharmacon) by using AMAXA nucleofection. Cells were cultured for 24h before stimulation with 100ng/ml LPS. Cells were lysed for protein extraction or RNA extraction.

The silencing of DynII or ATG12 in MCF-7 cells was conducted by transient transfection of Dharmacon smart pool siRNA oligos (final concentration 4 nM in 2 mL of a six well plate). For all siRNA experiments control cells (CTR) have been transfected with the same amount of transfection medium (RNAi Max+Optimem) but the siRNA oligonucleotide, as described elsewhere15 (link)16 (link)20 (link)46 (link). All other assays were performed as previously described15 (link)16 (link).

Transfection was carried out using Lipofectamine as described previously (Hjerpe et al., (link)
2012 (link)a,b). SMARTpool siRNA oligos from GE Dharmacon (Little Chalfont, UK) were used, siRNA against the DCNL1 3′UTR (5′-UACAUAGUCUGUACAAUAA-3′) was synthesized by Eurofins (Ebersberg, Germany).