Non targeting sirna 1

For depletion of PINK1 in HEK 293 cells, the following siRNAs were tested: PINK1 siRNA-1 (1199, Ambion, Grand Island, NY, U.S.A.), 5′-GGAGAUCCAGGCAAUUUUUTT-3′ and PINK1 siRNA-2, 5′-GACGCUGU UCCUCGUUAUGAA-3′ (S100287931, Qiagen, Venlo, Netherlands). Scrambled siRNAs with no known mammalian homology (non-targeting siRNA #1 (Ambion) and #2 (Santa Cruz Biotechnology, Dallas, TX, U.S.A.)) were used as negative controls. HEK cells were transfected with the siRNAs using the Mirus TransIT-TKO® reagent (Mirus, Madison, WI, U.S.A.) according to the manufacturer’s manual and then harvested after 48 h. Control vector-transfected cells were used as controls for all the experiments.

The following siRNAs were tested for depleting the indicated genes in the human primary fibroblasts or HEK cells: hspa9 siRNA (#1 and #2), acc1 siRNA (#1 and #2), fas siRNA (#1 and #2) (Ambion). Scrambled siRNA with no known mammalian homology (non-targeting siRNA #1 (Ambion) was used as negative control. Double siRNA treatment was performed by mixing two different siRNAs indicated at 1:1 ratio. Cells were transfected with the siRNAs using JetPrime according to the manufacturer’s manual and then harvested after 48 hr. Control vector-transfected cells were used as controls for all the experiments.