Cells were lysed in the existence of RNase inhibitors (Promega) and incubated with five mg precipitating antibody (mouse monoclonal anti-Assist antibody AIDA or mouse monoclonal anti-Flag antibody [Sigma]), followed by incubation with protein G agarose
The oligonucleotides had been annealed and ligated into the BamHI/EcorI lower pSIREN-ZsGreen according to OPC-8212the ``Knockout RNAi Methods User Manual'' from Clontech. The identical annealed shRNAs have been ligated into…