Knockdown of AMPKα1 in Hepatic Cancer

In order to reduce AMPKα1 protein expression in hepatic cancer cells, specific 21 nucleotide double stranded RNA (siRNA) and a scrambled control were designed as we previously described [37 (link)], and synthesized using an Ambion commercial kit SilencerTMsiRNA (Thermo Fisher Scientific). The following sequence, chosen according the guidelines described by Elbashir et al. [38 (link)], which targets AMPKα1 nucleotides 1842-1864 was used: AACATTTCTGCATATTAGGCTCCTGTCTC. Cells were transfected using Dharmafect 4 reagent (Thermo Fisher Scientific). Experiments were performed 24 h after transfection, and the specific decrease in AMPKα expression was confirmed by immunoblotting.