VRK1 Silencing Using siRNA Knockdown

Specific silencing of VRK1 was performed using two different siRNA from Dharmacon (DHARMACON RNA Technologies). The VRK1 sequences targeted by these siRNA oligonucleotides were siVRK1-02 (siV1-02), CAAGGAACCTGGTGTTGAA; and siVRK1-03 (siV1-03): That GGAAUGGAAAGUAGGAUUA. As negative control, the “ON-TARGETplus siCONTROL Non-targeting siRNA” from DHARMACON was used and is indicated as siCt in experiments. The efficiency of RNAi transfection was determined with “siGLO RISC-free siRNA” (DHARMACON). Briefly, cells were transfected with the indicated siRNA at a concentration of 20 nM using Lipofectamine 2000 Reagent (Invitrogen) or Lipotransfectin (Nivorlab) [18 (link),34 (link),74 (link)]. After transfection, cells were processed at the times indicated in specific experiments that were performed as previously reported [19 (link),77 (link)]. The depletion of VRK1 by siRNA has been previously reported for A549 [18 (link),21 (link),36 (link)] and HT144 [20 (link),21 (link)] cell lines.