Specific silencing of VRK1 was performed using siVRK1-02 (siV1-02), designed with the SMARTselection algorithm (Dharmacon, Lafayette, CO) and obtained from Dharmacon (DHARMACON RNA Technologies). The sequence target of siVRK1-02 is 5'-CAAGGAACCTGGTGTTGAA-3'. As negative control, the “ON-TARGETplus siCONTROL Non-targeting siRNA” from DHARMACON was used. The efficiency of RNAi transfection was determined with “siGLO RISC-free siRNA” (DHARMACON) labeled with a red fluorochrome [21 (link)].
Cells were transfected with the indicated siRNA at a concentration of 20 nM using Lipofectamine 2000 Reagent” (Invitrogen, Carlsbad, CA) according to manufactures instructions. After transfection, cells were processed for specific experiments as previously reported [14 (link), 17 (link), 24 (link)].
Cells were transfected with the indicated siRNA at a concentration of 20 nM using Lipofectamine 2000 Reagent” (Invitrogen, Carlsbad, CA) according to manufactures instructions. After transfection, cells were processed for specific experiments as previously reported [14 (link), 17 (link), 24 (link)].
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