VRK1 and Sox2 Knockdown in Cells

VRK1 knockdown was performed using three different siRNA for VRK1 (NM_003384): siVRK1-02 (siV1-02), siVRK1-03 (siV1-03) and siVRK1-09 (siV1-09) (Dharmacon RNA Technologies,). The sequences targeted by these VRK1 siRNA oligonucleotides were siVRK1-01: GAAAGAGAGTCCAGAAGTA; siVRK1-02: CAAGGAACCTGGTGTTGAA; si-VRK1-03: GGAAUGGAAAGUAGGAUUA; and siVRK1-09: AGGUGUACUUGGUAGAUUA. Sox2 was knockdown with the SOX2 Trilencer-27 human siRNA (Origene, Rockville, MD). As negative control the “ON-TARGETplus siCONTROL Non-targeting siRNA” (siCt) (Dharmacon) was used. The efficiency of RNAi transfection was determined with “siGLO RISC-free siRNA” (DHARMACON) labelled with a red fluorochrome. All of them have been previously used. Cells were transfected with the indicated siRNA (20 nM), using Lipofectamine 2000 (Invitrogen; Carlsbad, CA) as previously reported16 (link)17 (link)28 (link)38 (link)57 (link).